ANALYSIS CHART INCLUDING MICROFLUID SAMPLE PROCESSING STRUCTURE AND ELECTROCHEMICAL IMPEDANCE SPECTROSCOPY-BASED BIOSENSOR.

Abstract

The invention relates to a microfluidic analysis card for the reliable and reproducible determination of target analytes, such as mycotoxins, found in complex extracts obtained from food samples, by electrochemical impedance spectroscopy. The analysis card comprises an analysis cell (1), liquid inlet section (3), liquid outlet section (2), working electrode (5), counter electrode (6), reference electrode (7), magnetic elements carrying receptors (10), magnetic element (4) holding the magnetic elements in a specific and fixed pattern on the working electrode (5), flow constriction element (8) directing the liquid through the magnetic elements (10), bubble collection element (9) removing gas bubbles that may come into contact with the electrodes, and connection extensions (11) enabling the electrodes to be connected to an electrochemical reader.

Description

1 TARIFF MICROFLUID SAMPLE PROCESSING STRUCTURE AND ELECTROCHEMICALS IMPEDANCE SPECTROSCOPY-BASED BIOSENSOR-INCLUDED ANALYSIS CARD 5 Technical Area The invention relates to analytical measurement systems, microfluidic systems, and It is related to the technical fields of electrochemical biosensors. 10 More specifically, the invention involves a target within a liquid sample. for the detection and / or quantification of the analyte Specifically, processing the sample on a microfluidic card. electrochemical sampling with a sample processing structure together with an impedance spectroscopy-based biosensor 15 This relates to the analysis card containing the following. State of the Art Food safety, environmental analysis, biological analysis, clinical 20 In the fields of analysis and industrial process monitoring; a liquid rapid, precise and accurate analysis of target analytes within the sample electrochemical determination in a portable form Sensor systems are known. The target analyte in question is; a molecule, protein, antigen, antibody, microorganism, 25 cell, DNA, RNA, toxin, metabolite, chemical pollutant or It may contain other biological or chemical components. In known electrochemical sensor systems, most often a working electrode, a counter electrode and a reference electrode 30 Electrode assemblies containing electrodes are used. Electrodes offer cost advantages and suitability for mass production. to provide a plastic, ceramic or similar carrier 2 screen-printing method on the substrate can be created. The working electrode surface is placed on the target Antibodies and antigens that can selectively interact with the analyte. aptamer, enzyme, nucleic acid probe, or similar receptor The structures are immobilized; the target analyte and receptor 5 electrochemical formation resulting from the interaction between them The change is being measured. One of the electrochemical measurement techniques In electrochemical impedance spectroscopy, the electrode and 10 impedance occurring at the interface between the measuring fluid The changes are evaluated. Thus, the receptor's function is assessed. its presence on the electrode, the target analyte to the receptor bonding, washing processes or other on the electrode surface Using the signal difference resulting from the changes, 15 The presence and / or quantity of the analyte can be determined. EIS-based. In biosensors, a reference impedance value is used before measurement. re-uptake and analyte-receptor interaction after Measurement is a known practice. However, especially food, soil, environmental fluid, complex biological fluid and process samples The samples contain numerous components in addition to the target analyte. Fat, protein, carbohydrate, salt, pigment, particle, cell residue, organic component, non-microorganism biological 25 components and similar components are adsorbed onto the electrode surface. It can clog microfluidic lines, electrochemically it can interfere with the signal or the identification of the target analyte This can create a matrix effect that makes analysis more difficult. Therefore, analysis homogenization of the sample beforehand, extraction, 30 dilution, filtration, separation, enrichment, washing and / or appropriate analysis It may need to be conditioned with the solution. 3 For this purpose, a device that carries and directs the sample in small volumes, microfluid that separates and subjects to reaction stages Microfluidic systems are used; sample inlets, reagent inlets, microchannels, mixing sections, incubation chambers, filtration / separation structures, 5 These may include valves, waste containers, and sensor areas. By arranging structures on a chip or card, analysis the process is portable, more controlled and more user-friendly The aim is to make them less dependent. US2023264192A1 number and “Microfluidic Electrochemical The patent document titled "Analyte Detectors" deals with microfluidics. It is aimed at electrochemical analyte detection on chips. The document discusses small molecules and macromolecules, for example. Microfluidic chip 15 that can be used in the detection of analytes structures; electrochemical sensor sections, flow channels and the integration approach with the analysis device is explained. The document discusses point-of-care applications of microfluidic chips. its use and microfluidic structure and electrochemical 20 regarding bringing the measurements together on the same platform It is understood that this is the case. The aforementioned document discusses microfluidic systems and electrochemical processes. in terms of integrating the analyte detection component This constitutes a significant example of the current technique. With this 25 together, general purpose microfluidic-electrochemical analysis in its approach, to extracts obtained from food samples elements of the specific complex matrix affecting EIS measurement reduction; for this purpose, physical and chemical decomposition, incubation, washing and conditioning of the analysis medium 30 processes as a whole in accordance with EIS measurement The design is also a technical matter that needs to be solved. This emerges as a problem. 4 In other words, it is simply a microfluidic channel structure. and the electrochemical sensor are located within the same device, Reliable EIS in complex samples such as food extracts It may not be sufficient for measurement. Especially the sample. The interfering components it contains have a 5 on the electrode surface. It masks receptor-analyte interaction, serving as both reference and measurement. distortion of the difference between signals, non-on electrode surface forming specific bonds or in flow elements There are technical issues such as the accumulation of waste. Therefore, the microfluidic structure has only one function: sample transport. 10 no, in a way that will increase the reliability of EIS measurement structuring the sample with processing and conditioning functions is required. EP4180805B1 numbered and “Screen-printed electrode, 15 manufacturing method thereof, electrochemical sensor consisting of said electrode for detecting water pollutants, Patent document titled "and operating method of said sensor"; screen- for detecting pollutants in water samples printed electrode structure, this electrode's production method and 20 It describes its use in electrochemical sensors. The document describes a study conducted on a carrier. containing electrode, counter electrode and reference electrode with a screen-printed electrode structure Solutions for use in pollutant detection are included in section 25. It is receiving. This document can be low-cost and disposable. screen-printed electrodes in electrochemical analyses This shows that it can be used. However, this approach, 30 essentially depends on the structure of the electrode, its production, and the water. It focuses on the detection of contaminants in food samples. formed as a result of grinding and extraction of samples high matrix effect fluids, in a microfluidic card pre-processing; then electrochemical impedance a biosensor measurement suitable for spectroscopy a multi-stage process such as directing to the region not accepted. 5 In known SPE solutions, especially for complex foods direct contact of the extracts with the electrode; electrode surface contamination, non-receptor-specific the formation of interactions, signal-reproducibility 10 problems arising from different types of food This can lead to variable matrix effects. Furthermore, classical In drop-in strip or electrode assemblies, the reference the measurement solution, the sample containing the target analyte, washing controlled solution and, if necessary, measuring solution, 15 repeatable and isolated measurements It is difficult to deliver it to the region. Within the known technical framework described above, the existing 20 solutions contain the following technical problems: is being evaluated:  In extracts obtained from complex food samples Interfering components found in electrochemical measurement It creates a matrix effect and the EIS signal is 25 This negatively affects its credibility.  In microfluidic-electrochemical analysis systems sensors and flow lines being on the same platform Although it is known, the food extract must be made suitable for EIS measurement. 30 physical and / or chemical requirements for delivery Separation, washing, incubation, and conditioning 6 steps reliably as an on-card process It needs to be managed.  Screen-printed electrodes offer low cost and mass production. while providing an advantage, especially in complex situations 5 non-specific electrode surface in samples susceptible to interactions, contamination, and signal distortions. This means that reliable, repeatable, and quantitative results can be obtained. This makes it difficult to do.  In classic electrode / strip systems, the sample dripping, incubation, washing such as performing and applying the analysis solution The processes depend significantly on the user application. This situation leads to variability between analyses and field 15 This leads to a lack of standardization in its use.  Different receptor structures for different types of analytes Although they can be used, different versions of the same platform 20 to sample types or target analytes a modular and controlled system that will allow it to be adapted A card architecture is needed. In this context, complex liquids, especially food samples. The samples were subjected to EIS measurement in a microfluidic structure for 25 days. proper processing; screen-printed electrode based and impedance resulting from receptor-analyte interaction repeatable detection of change An analysis card providing this information is needed. 30 7 Problems that the invention aims to solve. The main purpose of the invention is to analyze food samples in particular. complex liquid containing a high percentage of interfering components The electrochemical impedance of the target analytes in the samples is 5. reliable, repeatable and spectroscopic spectroscopy method to be determined in as standardized a way as possible to provide. 10 based on electrochemical impedance spectroscopy in biosensors, located on the surface of the working electrode Specific binding between receptor structure and target analyte as a result, the electrical properties of the electrode-liquid interface is changing. This change is related to the reference measurement. 15 from the interaction of the analyte-containing sample with the receptor surface then through the impedance difference between the measurements obtained It can be determined. However, food extracts and similar products... Complex samples; containing protein, fat, in addition to the target analyte, carbohydrate, pigment, salt, particle, cellular debris, different organic components and other electrochemically 20 They may contain active or passive substances. These components, It can disrupt analyte-receptor interaction, electrode It can cause non-specific bonding on its surface and EIS By modifying the measurement signal, the accuracy of the analysis can be improved. It can lower it. 25 Therefore, the main technical solution that the invention aims to provide The problem arises in food or similar complex samples. reducing matrix effect and targeting the analyte in EIS measurement. 30 in the electrochemical biosensor region under suitable conditions The goal is to achieve control over the matrix effect. If this is not possible, then the same target analyte concentration should be used. Different samples have different impedance results. 8 can be obtained; false positive, false negative or Quantitatively inaccurate results may occur. Another technical problem that the invention aims to solve is the sample. 5 preparation and analysis phases in user application It is addictive. Known as drip or dip. In screen-printed electrode sensors, the sample adheres to the electrode. application, storage, washing, reagent / analysis such as applying the solution and starting the measurement. The steps vary in duration, volume, and number by different users, and are 10... This can be achieved under the application conditions. This situation, especially on-site analysis and routine quality control In these applications, it causes variability among measurement results. is happening. In this context, the invention relates to a sample on an analysis card. processing within a microfluidic structure by following a specific path It aims to meet the need. The sample, analysis physical and / or chemical after being recorded on the card Separation, filtration, dilution, reaction, incubation, 20 washing, solution change and / or conditioning being subjected to one or more of the processes; target the analyte and / or the medium that will interact with the target analyte The aim is to make it suitable for EIS measurement. An additional problem that the invention aims to solve is microfluidics. the system and the electrochemical sensor on the same board on platforms where it is known to be present, especially those with complex matrices The goal is to ensure analytical reliability of the samples. A 30 of microfluidic channels and electrochemical sensors simply bringing them together; the electrode surface contamination, non-specific binding, sample components accumulation of impedance signal in the measurement region 9 It may not prevent it from being affected by initiatives. General purpose. microfluidic-electrochemical analyte detection systems, electrochemical sensing can be performed on a microfluidic chip. However, complex methods like food extracts are involved. 5 regarding the specific conditioning of samples for EIS measurement. Technical needs persist. Similarly, screen-printed electrodes are low-cost, portable and suitable for mass production electrochemical Although it is a known approach to use in analysis, 10 these electrodes directly interact with complex food extracts. measurement reliability and repeatability in contact Problems may arise with the screen-printed electrode. The study involves determining the counter and reference electrode arrangement. controlled, sufficient application to the electrode area alone 15 a cleaned sample / measurement solution suitable for EIS measurement This does not mean that it is provided. Therefore, SPE-based the sample preparation and conditioning function of the electrode structure functionally with visible microfluidic board sections They need to be combined. 20 The invention is also adaptable to multiple types of analytes. It aims to solve the problem of providing analytical infrastructure. Target analytes in food safety applications include pathogens, toxins, allergen, protein, small molecule, nucleic acid or chemical 25 It may be residual in nature. However, it is the same basic principle. problem, environmental sample, water sample, soil extract, also in different fields such as biological fluid or process sampling This is revealed. Therefore, the functioning of different receptors... will allow its use on the electrode, this 30 However, sample processing and EIS measurement conditions were controlled. A card infrastructure is needed to manage this in this way. In conclusion, the invention demonstrates the matrix effect in complex samples. reducing, suitable sample conditions for EIS measurement ensuring standardization of sample preparation and measurement workflow. to reduce interference on the electrode surface, User-generated implementation differences should be minimized by 5. downloadable and adaptable to different analytical types problems of creating an electrochemical analysis platform It aims to solve the problem. Explaining the Figures 10 Figure 1. Perspective view of the analysis cell. Figure 2. Side view of the analysis cell. Figure 3. Perspective cross-sectional view of the analysis cell. Figure 4. Structural view of the electrodes, 15 Figure 5. Appearance of the system consisting of electrodes. Figure 6. The system consisting of electrodes on a magnet. appearance, Figure 7. Magnet parallel to magnetic particles. the appearance of the structure in which it is placed, 20 Figure 8. Side view of the system. Explanation of References in Figures 1. Analysis cell 2. Liquid outlet section 25 3. Liquid inlet section 4. Magnetic element 5. Working electrode 6. Counter electrode 7. Reference electrode 30 8. Flow restriction element 9. Bubble collecting element 10. Magnetic elements 11 11. Link extension Disclosure of the Invention The invention describes the process of detecting a target analyte in complex liquid samples, 5 especially in extracts obtained from food samples the mycotoxins found were analyzed by electrochemical methods. microfluidic analysis card and analysis for the purpose of It is related to cell (1). In the system covered by the invention, the target analytics are specifically designed for this purpose. receptors that can bind to magnetic particles They are immobilized. Receptors; aptamer, antibody, antibody fragment, antigen, enzyme, nucleic acid probe, molecular 15 with printed polymer, peptide, bacteriophage or target analyte There may be another receptor that can selectively interact with it. One In practice, the receptor is a single-stranded DNA structure. It is an aptamer. A receptor-functionalized magnetic field. The particles are placed on the analysis card in a dry state, preferably frozen. dried, or in other words, lyophilized form 20 They can be positioned. Thus, the receptors are stored during this process. to maintain its functionality, extend the shelf life of the card, and receptor activation required before analysis It is ensured that it is carried out in a controlled manner. An analysis card in one application of the invention; an analysis cell. (1) is in fluid communication with the analysis cell (1). liquid inlet section (3), a liquid outlet section (2), working electrode (5), counter electrode (6), reference electrode (7), magnetic element (4), flow constriction element (8), bubble 30 collecting element (9), magnetic elements (10) and electrodes a connector that allows connection to an external analysis device includes extensions (11). 12 The analysis cell (1) holds the liquid sample, the rehydration solution, controlled washing solution and / or analysis solution It is the microfluidic measurement volume to which it is brought into contact. The analyzer cell (1) has one input end to the other output end. narrower in the direction of flow, preferably in the inlet and outlet areas. 5 and will have a larger interior volume in the central area It can be shaped. Thanks to this geometry, the analysis of the liquid can be done. more magnetic particles located in the center of the cell (1) effective contact is ensured. The analysis cell (1) in general 10 with a narrow entrance and exit, widening towards the middle. It can be in one form. However, the analysis cell (1); elliptical, oval, circular, polygonal, rectangular, conical, prismatic or flow to the reaction area in the center It may also have another geometry to guide it. Liquid inlet section (3) into the analysis cell (1) rehydration solution, primary washing solution, the second washing solution, analysis containing redox probe of the solution and / or sample solution containing the target analyte It enables the intake of liquid inlet section (3), microfluidic 20 the card has an input port, connection channel, hose connection, in the form of a reactive input port or a combination thereof It can be formed. Liquid inlet section (3), one or more It can be connected to the liquid line. These liquid lines are connected to the analysis board. a syringe pump in an external analysis device, 25 vacuum pump, pressure source, peristaltic pump, valve control by system or similar flow control elements It can be done. Liquid outlet section (2) passes through the analysis cell (1) 30 the liquid is recycled into a waste container after analysis. to the production line, sample packaging volume, or for reuse It directs the liquid to the inlet section (3). Liquid 13 The output section (2) is performed inside the analysis cell (1). a rehydration, flushing, reference measurement, or analyte binding After this step, it is possible to remove the liquid used. In one application, the liquid outlet section (2) provides the target analyte. forward through the analysis cell (1) of the liquid package containing 5 a reverse that allows it to be passed through repeatedly in the reverse direction It can be associated with the circulation line. Thus, the target the analyte located on magnetic particles probability of binding to receptors and binding efficiency can be increased. 10 The working electrode (5) is placed inside the analysis cell (1) or the analysis on a surface that forms a boundary with cell (1) is positioned. The working electrode (5), preferably It has a rectangular or elongated geometry. 15 However, the working electrode (5); strip, square, circular, oval, comb-shaped, interdigitated, polygonal or magnetic particles regularly in the measurement area It may have a different geometry that makes it possible to hold it. Working electrode (5), carbon, graphite, gold, platinum, silver, 20 conductive polymer, metal oxide, carbon nanotube, graphene, metal nanoparticles or combinations thereof can be created. In one application, the working electrode (5), a plastic, ceramic, polymer by screen-printing method It is formed on a film or other carrier surface. 25 Counter electrode (6) and reference electrode (7), study three electrode electrochemical electrode (5) Analyzer cell (1) to form the measurement setup inside or on a surface adjacent to the analysis cell 30 is positioned. The counter electrode (6) is electrochemical It contributes to the completion of the circuit. Reference electrode (7) the electrochemical potential relative to a stable reference 14 It enables monitoring. Reference with counter electrode (6) electrode (7), near working electrode (5) and analysis will make simultaneous contact with the fluid inside the cell (1) It is positioned in this way. Thus, the electrochemical impedance during spectroscopy, the working electrode (5) on 5 impedance resulting from receptor-analyte interaction Changes can be detected more consistently. Reference electrode (7), for example a silver / silver chloride based electrode it could be. Magnetic element (4) is placed under, opposite the working electrode (5), magnetic adjacent to or working electrode (5) another that will create a magnetic field on the particles It is the magnetic structure placed in position. Magnetic element (4), permanent magnet, electromagnet, permanent magnet, neodymium 15 magnet, ferrite magnet, electromagnetic coil or It may include combinations of these. In an application magnetic element (4), under the working electrode (5) It can be installed in the space in a removable and reattachable manner. Thus, from the production, storage or analysis of the card 20 effectiveness of magnetic element (4) before / after stage It may be adjustable or detachable from the card. Magnetic element (4), preferably along the working electrode (5) 25 that extends and aligns with the area under the working electrode (5) It creates a longitudinal magnetic field. In one example, When the working electrode (5) is rectangular, the magnetic element (4) working electrode (5) extending along the length and from above as a magnet with a triangular cross-section in appearance It can be formed. A triangular cross-section magnetic element (4) of 30 the edge parallel to the area under the working electrode (5) or They are positioned adjacent to it. In this way, magnetic particles, Instead of randomly clustering on the working electrode (5), a strip or line extending along the length of the electrode It can be maintained in this state. Such a placement is based on a reference measurement. despite the subsequent flow and washing processes to preserve the spatial distribution of magnetic particles and It contributes to increasing the repeatability of measurements. 5 Magnetic elements (10) that can bind specifically to the target analyte. ferromagnetic, ferrimagnetic or receptor-carrying They are superparamagnetic particles. Magnetic elements (10), iron oxide, magnetite, maghemite, ferrite, metal-coated 10 polymeric particles or similar magnetic properties Magnetic elements (10) can be produced from materials, spherical, hemispherical, rod, disc, scale, fiber, microbead, nanobead or in similar forms. Magnetic elements (10) immobilized 15 target analyse-specific receptors by binding between the target analyte and the receptor the reaction is constant in relation to the working electrode (5) This can be performed in the reaction zone. Magnetic elements (10) inside the analyzer cell (1) 20 on the working electrode (5) or working electrode (5) It is positioned between the fluid flow path and the fluid flow path. Magnetic element (4), magnetic elements (10) working It keeps the electrode (5) fixed on the reference electrode. Thus, it provides a reference electrode (5). Fluid passage, flushing, forward-25 occurring after EIS measurement. reverse flow and / or repeated sample packing magnetic elements (10) significantly during circulation This prevents displacement. This situation affects the reference measurement. magnetic element distribution after analyte binding unwanted 30 between magnetic element distribution in the measurement It helps to reduce variations. Accordingly, the measured impedance difference is predominantly analyte-receptor The aim is for it to result from interaction. 16 Flow constriction element (8), magnetic inside the analyzer cell (1) local fluid flow cross-section in the region where elements (10) are located It is a structure that reduces flow. Flow constriction element (8), analysis a projection extending downwards from the upper surface of the cell (1), roof extension, fin, barrier, bottleneck, partial closure 5 element, plate or other flow cross-section reduction element It can be created as follows. Flow restriction element (8), working held on electrode (5) and working electrode (5) magnetic elements (10) on or near this area It is located at position 10. Thanks to the flow constriction element (8), the sample solution forced to pass through magnetic elements (10) This ensures that the target analyte in the liquid reaches the receptor. The probability of contact with magnetic elements (10) is 15 The middle region of the analysis cell (1) is raised. expanded volume and magnetic elements (10) flow narrowing element (8) that reduces the flow cross-section in the region By using them together, on the one hand, the magnetic field of the liquid... effectively passing through the (10) region of the elements, the other 20 On the other hand, the magnetic field of the fluid flow velocity in the region in question causing the elements (10) to change position due to the flow effect The aim is to control it to an extent that it will not happen. Multiple streams within the analysis cell (1) in an application 25 A constriction element (8) may be included. For example, the working electrode (5) three flow restriction elements spaced apart along (8) can be created. This arrangement involves magnetic elements in the liquid. Instead of quickly passing through a single point (10), different the orientation of the regions and the target analyte and receptors 30 This can enable a higher level of interaction between them. Flow restriction elements (8), fixed, adjustable, elastic, These can be flexible, movable, or valve-functioning structures. 17 Bubble collection element (9) inside the analysis cell (1) separated from the liquid or entering the system during liquid transfer limiting contact between gas / air bubbles and electrodes a volume, pocket, space, cavity or arranged in such a way It is the guiding channel. Bubble collecting element (9), liquid 5 near the inlet section (3) and / or liquid outlet section (2), preferably working electrode (5), counter electrode (6) and The reference electrode (7) is positioned away from the region. In an application, the bubble collecting element (9) is a collapsed 10 widening or sloping upwards starting from the region a channel rising in this way, a bubble escape route and its It contains a pocket of volume located at the end. Air / gas within the liquid. bubbles, due to density difference and fluid dynamics It tends to move upwards. Bubble collection 15 The inclined and rising structure of element (9), bubbles collecting bubbles by removing them from the electrode surface It directs the bubbles towards element (9). working electrode (5), counter electrode (6) or reference the electrode (7) should remain on the electrode; the electrode-liquid contact area should be 20 because it can alter and cause unwanted deviations in the EIS signal. because it can cause bubble collection element (9) measurement It is important in terms of accuracy and repeatability. Connection extension (11), working electrode (5), opposing 25 electrode (6) and reference electrode (7) a potentiostat, with reader device, measuring circuit or external control unit a conductive line that enables an electrical connection, contact pad, terminal, socket, connector, connecting strip or It is a similar electrical connection structure. Connection extensions 30 (11), on one side of the analysis card, on its bottom surface or a link located away from the analysis cell (1) It can be located in that region. Thus, the analysis card is an analysis. 18 by plugging it into the device or connecting it to an external reading module electrochemical impedance spectroscopy measurement It makes it possible to carry it out. Working Principle 5 The analysis process covered by the invention is as follows in an application: This is carried out in sequence. First, a magnetic field containing a receptor is used. elements (10), working electrode inside analysis cell (1) (5) or liquid flow path with working electrode (5) 10 It is located between them. Magnetic element (4), magnetic elements (10) on the working electrode (5) constants in a given distribution. 15 Lyophilized receptor-carrying magnetic elements (10) analysis via liquid inlet section (3) for activation Rehydration solution is given to the cell (1). Rehydration solution, receptors on magnetic elements (10) suitable for rehydration and binding of the target analyte. It enables it to become active. Rehydration used 20 The solution can be removed from the liquid outlet section (2) or if necessary, within the analysis cell (1) for a certain period of time It can be held. After the rehydration process, the fluid inlet section (3) 25 first washing solution through the analysis cell (1) can be given. First washing solution; undissolved residues, free receptor components, or removing other unwanted elements from the analysis It helps. First washing solution, liquid outlet 30 from section (2) to the waste line or waste container is directed. 19 Afterwards, the analysis solution containing the redox probe was analyzed. The analysis solution is taken into the cell (1). The working electrode (5), counter electrode (6), reference electrode (7) and magnetic The elements (10) are brought into contact with the connection extensions (11). potentiostat or analyzer 5 to which the card is connected The first EIS measurement is performed. This first measurement is the target. Magnetic elements carrying analyte-unbound receptors (10) It establishes the reference or base impedance value. After the reference measurement, the analysis cell (1), first wash 10 with solution or another suitable washing solution It can be cleaned. Thus, it is used for reference analysis. The analysis solution and any free components are removed. At this stage, the magnetic element (4), magnetic elements (10) It maintains its position on the working electrode (5). Flow 15 constriction element (8), flow to magnetic elements (10) while directing the flow, the magnetic elements (10) by dragging it to change its distribution in the reference measurement boundaries. The bubble collecting element (9) is flow-borne. or gas produced during liquid transfer 20 It removes the bubbles from the electrodes. Sample solution containing the target analyte, especially for food. The extract prepared from the sample, after the necessary preliminary processes then through the liquid inlet section (3) into the analysis cell 25 (1) is taken. In one application, the sample solution is taken into the analysis card. chemical found in another microfluidic section It is passed through a purification / separation column. Chemical purification / separation column; oil from food matrix sources, Reduction of protein and similar interfering components 30 for this purpose C18, PSA or adsorbent suitable for the sample type and / or It may contain a separating material. This allows the target analyte to be contained within. conditioning of the solution before electrochemical measurement and This helps to reduce the matrix effect. Purified target analyte solution, in an application They are divided into liquid packages of specific volumes. The liquid packages are 5 air transfer, positive pressure, negative pressure, syringe pump, vacuum pump or a combination thereof It is sent into the analysis cell (1) via each liquid pack, from the region where the flow constriction element (8) is located by passing through the magnetic elements (10) located on 10 They are brought into contact with receptors. Liquid packets are moved forward or backward inside the analysis cell (1). It can be moved in one direction or in back-and-forth loops. In one application, the liquid package, liquid inlet section (3) and liquid 15 a certain number of back-and-forth movements between exit section (2) by making magnetic elements (10) in the region where they are located It is passed through repeatedly. In another application, a liquid pack. inside the analysis cell (1) via a return circulation pathway It is circulated again. This repeated pass involves 20 passes of the target analyte. It increases the number of contacts with the receptor and the probability of binding. Moving the liquid package in specific volumes, diffusion and concentration in high-volume sample flow low binding efficiency that may arise due to the decrease It helps to reduce it. 25 The target analyte is located on the magnetic elements (10). After binding to the receptors, the sample solution used through the liquid outlet section (2) to the waste tank or waste is sent to the pipeline. For multiple liquid packages, this process takes 30 minutes. It is repeatable. Thus, in a high volume sample solution. external and pre-concentration of the target analyte found. no need for expensive column systems or long waiting times 21 without sensing, by receptors with higher efficiency The aim is to capture him. After the sample binding stage is complete, the analysis cell (1) It can be washed with the second washing solution. Second washing 5 the solution contains sample components that are not bound to the target analyte, residues originating from the food matrix and the measurement signal to reduce other components that could have a negative impact It may contain components. Then the first wash. An additional wash can be performed with the solution. Thus, 10 Target analyte–receptor complexes magnetic elements (10) while held on, non-specifically attached or The free-floating components are removed. In the final stage, the analysis solution containing the redox probe is again 15 It is sent to the analysis cell (1). Working electrode (5), using counter electrode (6) and reference electrode (7) A second EIS measurement is performed. After analyte binding. analyte binding with the next second impedance value The previous reference impedance value is compared. These measurements are 20. the impedance difference between them, for example, in the load transfer resistance change, change in impedance modulus, change in phase angle change, capacitance change or equivalent circuit It can be determined through at least one of its parameters. The determined difference depends on the presence of the target analyte and / or 25 It is used to produce results regarding concentration. Alternative Application In another application of the invention, the analysis cell (1), 30 instead of being created on a planar card surface in the form of a layered and / or three-dimensional structure can be created. In this application, the working electrode (5) is the opposite 22 electrode (6) and reference electrode (7) are different from each other at different height levels or side by side on the same base It can be positioned. Magnetic element (4), working while being placed under the electrode (5); flow restriction elements (8), three 5 in the measurement region where the electrodes are located It can be located at one or more points. Bubble collection elements (9) are located on the sides of the measurement area, entrance and exit multiple located in the sections or away from the electrodes It can be arranged in a bubble pocket shape. In another application, the input and output lines of the analyzer cell (1) It forms a closed flow circuit with the liquid inlet section (3) and liquid outlet section (2), analysis cell of the same liquid package (1) so that it can be passed through more than once It is connected to a recirculation structure. Thus, the liquid sample, 15 magnetic elements before electrochemical analysis (10) It is passed through in multiple cycles. In another application, the magnetic element (4) is a permanent magnet. 20 can be checked instead of or in addition to this. It is created in the form of an electromagnet. The electromagnet magnetic field generated; rehydration, reference measurement, from sample bonding, washing or final EIS measurement steps By changing at least one of the magnetic elements (10) the operation The holding force on electrode (5) can be adjusted. 25 In another application, the target analyte is examined in food samples. The mycotoxin found is aflatoxin. However, the invention also includes aflatoxin, ochratoxin, deoxynivalenol, zearalenone, fumonisin, patulin, pesticide, antibiotic residue, veterinary drug residue, 30 allergen, toxin, microorganism, virus, bacteria, protein, DNA, RNA, small molecules, or other similar target analytes It can also be used in detection. The application area of ​​the invention is food. 23 not limited to safety; environmental analysis, water analysis, soil analysis, clinical analysis, biological fluid analysis and It can also cover areas such as industrial process analysis.

Claims

24 REQUESTS 1. Electrochemical detection of a target analyte. It is a microfluidic analysis chart for this purpose; an analysis cell (1), 5 in fluid communication with the said analysis cell (1) at least one liquid inlet section (3) and at least one liquid outlet section (2), a study located in analysis cell (1) electrode (5), a counter electrode (6) and a reference electrode 10 electrode (7), at least one that is suitable for selective interaction with the target analyte. containing the receptor and on the working electrode (5) multiple magnetic elements arranged in such a way as to be held man (10), 15 magnetic elements (10), working electrode (5) on- working in a way that holds it in a defined position. to form a magnetic interaction with the electrode (5) position the working electrode (5) under and / or near it. at least one magnetic element (4), 20 inside the analysis cell (1), magnetic elements (10) It also locally affects the fluid flow cross-section in the region where it is located. by passing the liquid through the magnetic elements (10) at least one flow restriction element to direct your urine (8), formed in the analysis cell (1) and / or liquid ingress 25 transported between section (3) and liquid outlet section (2) gas bubbles working electrode (5), opposing electrode- will move the electrode (6) away from the reference electrode (7) at least one bubble collecting element arranged in clay (9) and 30 working electrode (5), counter electrode (6) and reference electrode (7) to an external electrochemical measuring device at least one arranged to be connected electrically link extension (11) microfluidic analysis characterized by its inclusion card.

2. Microfluidic analysis chart according to claim 1, analysis 5 liquid inlet section (3) of cell (1), liquid outlet in a flow direction extending toward section (2), the entrance and a wider central area compared to the exit sections microfluidic analysis characterized by its inclusion card. 10 3. This is a microfluidic analysis chart according to claim 2, and the work is as follows: analysis of the electrode (5) and magnetic elements (10), located in the central region of the cell (1) microfluidic analysis characterized by its gradual reduction card. 15 4. Microfluidic according to any of claims 1 to 3. analysis card, magnetic element (4), working elect- positioned under the rod (5) and working elect- magnetic field along the longitudinal direction of the rod (5) The KA-20 contains a magnet that extends in such a way as to create a magnetic field. Characterized microfluidic analysis card.

5. Microfluidic analyzer card according to claim 4, magnetic. element (4) directed under the working electrode (5) a magnet with a triangular cross-section having at least one side Microfluidic analysis chart characterized by its... 25 6. Microfluidic analysis chart according to claim 4 or 5, magnetic element (4), magnetic elements (10) working in a longitudinal strip on the electrode (5) characterized by its structure designed to hold Microfluidic analysis chart. 30 7. Microfluidic according to any of claims 1 to 6. analysis card, magnetic elements (10), iron arrow- sit, magnetite, maghemite, ferrite, metal-coated polymeric 26 by containing particles or a combination thereof Characterized microfluidic analysis card.

8. Microfluidic according to any of claims 1 to 7. analysis card, magnetic elements (10), analysis in its cell (1) dry and / or lyophilized state-5 microfluidic analysis card characterized by its properties.

9. This is a microfluidic analysis chart according to claim 8, and the aforementioned... Nusu receptor; aptamer, antibody, antibody fragment, antigen, enzyme, nucleic acid probe, molecular imprint being at least one of the following: polymer, peptide or bacteriophage-10 microfluidic analysis card characterized by its properties.

10. Microfluidic according to any of claims 1 to 9. analysis card and flow restriction element (8), analysis from the upper surface of the cell (1) to the working electrode (5) 15 which extends correctly and contains magnetic elements (10) a protrusion, ceiling that reduces the flow cross-section in the area extension, barrier, plate and / or partial closure element microfluidic analysis card characterized by its...

11. Microfluidic analysis chart according to claim 10, main- Working electrode (5) bo-20 inside lys cell (1) several spaced apart characterized by having a flow constriction element (8) microfluidic analysis chart.

12. Microfluidic according to any of claims 1 to 11. analysis card and bubble collection element (9), main-25 up through the fluid flow path inside the lysate cell (1) an extending sloping bubble guiding channel and word the subject is a balloon located at the end of the routing channel. microfluid characterized by containing a pocket. Analysis card. 30 13. Microfluidic analysis chart according to claim 12, bubble collection element (9), from the working electrode (5), from the counter electrode (6) and the reference electrode (7) 27 characterized by its location in a remote area. len microfluidic analysis card.

14. Microfluidic according to any of claims 1 to 13. analysis card and working electrode (5), opposite electrode (6) and reference electrode (7), a carrier 5 Created on a substrate using screen-printing method. microfluidic analysis card characterized by its...

15. Microfluidic analysis chart according to claim 14, and... working electrode (5) carbon, graphite, gold, platinum, silver, conductive polymer, carbon nanotube, graphene, metal 10 contains nanoparticles and / or combinations thereof. Microfluidic analysis card characterized by its properties.

16. Microfluidic analysis chart according to claim 14 or 15. and the reference electrode (7) silver / silver chloride microfluidic analysis characterized by its inclusion 15 card.

17. Microfluidic according to any of claims 1 to 16. analysis card and liquid inlet section (3), a rehydr- ration solution, at least one washing solution, target 20 containing sample solution containing analyte and redox probe At least two of the analysis solutions are placed in the analysis cell. (1) structured in such a way as to allow selective delivery. microfluidic analysis characterized by its classification card.

18. Microfluidic 25 according to any of claims 1 to 17. It is an analysis card, with liquid inlet section (3) and liquid outlet. section (2), main of the liquid package containing the target analyte. from the lysate cell (1) multiple forward and / or backward directions a return circulation that will allow it to be passed through several times 30 characterized by being in fluid communication with the flow line. microfluidic analysis card obtained.

19. Microfluidic according to any of claims 1 to 18. analysis card, before entering the analysis cell (1) 28 to condition the liquid sample containing the target analyte structured, containing adsorbent and / or separator material. It is characterized by containing a chemical purification section. Microfluidic analysis card.

20. Microfluidic analysis chart according to claim 19, Kim-5 legal purification section, C18, PSA, solid phase extract- reaction material, adsorbent, ion exchange material, immunoaffinity material or combination thereof. micro- characterized by containing at least one of them Fluid analysis chart. 10 21. Microfluidic analysis chart according to claim 19 or 20. and located at the exit of the chemical purification section and the next flow stage of the liquid containing the target analyte- selectively blocking or allowing exam access 15 including a flexible flow pipe structured accordingly characterized by having a valve interface microfluidic analysis chart.

22. Microfluidic analysis card according to claim 21, valve the interface, located outside the analysis card -20 made to be compressed by the solenoid unit It is characterized by containing a pinch valve section that has been folded down. microfluidic analysis card obtained.

23. Microflow according to any of claims 19 to 22. blood analysis card, in the chemical purification department an outlet used to direct the liquid through a vacuum 25 point and at the said exit point the liquid has a va- It contains a hydrophobic membrane that prevents it from passing into the sand line. Microfluidic analysis card characterized by its properties.

24. Microfluidic according to any of claims 1 to 23. This is an analysis card, and the liquid containing the target analyte is placed in a specific 30-minute interval. (1) in volumes of liquid packages into the analysis cell It is characterized by containing an analyte chamber for delivery. Microfluidic analysis card. 29 25. Microfluidic according to any of claims 1 to 24. analysis card, connection extensions (11), working electrode (5), counter electrode (6) and reference electrode rod (7) a potentiostat and / or electrochemical em- 5 with a reader device that will perform pessimism spectroscopy including contact pads for electrical connection. microfluidic analysis card characterized by its properties.

26. Microfluidic analysis chart according to claim 25, main- magnetic receptor found in the lys cell (1) elements (10) before interacting with the target analyte and he-10 impedance measurement taken after interaction with the analyte. in a way that will allow comparison of them microfluidic characterized by its structuring Analysis card.

27. Microfluidic 15 according to any of claims 1 to 26. This is an analysis chart, and the target analyte is mycotoxin, aflatoxin- sin, ochratoxin, deoxynivalenol, zearalenone, fumonisin, patulin, pesticide, veterinary drug residue, antibiotic- tick residue, allergen, microorganism, virus, bacteria, protein, DNA, RNA or at least one of the small molecules - 20 Microfluidic analysis card characterized by its properties.

28. Microfluidic according to any of claims 1 to 27. It is an analysis card, and the card contains data obtained from a food sample. an extract, water sample, soil extract, environmental target 25 in sample, biological fluid or process sample characterized by being geared towards the detection of the analyte microfluidic analysis chart.

29. Electron sequencing of the target analyte in a liquid sample. It is a method for the chemical detection of... edi-30 is characterized by including the following process steps: Len method: an analysis of magnetic elements containing receptors (10) working electrode (5) located inside cell (1) magnetic interaction with the working electrode (5) on it retained through the magnetic element (4) that forms, magnetic elements (10), from the liquid inlet section (3) a rehydration solution transferred to the analysis cell (1) Activation with 5 working electrode (5), counter electrode (6) and reference Using the electrode (7), magnetic containing the receptor before the elements (10) interact with the target analyte reference electrochemical impedance spectroscopy measurement- the realization of the project, 10 the liquid sample containing the target analyte into the analysis cell (1) transmission and analysis cell (1) magnetic Passing through the area where the elements (10) are located magnetic elements in the liquid sample containing the target analyte Binding of target analyte to receptors on (10)-15 In order to provide the class, further inside the analysis cell (1) multiple in one direction, in the reverse direction and / or in forward-backward direction moved once, from the analysis cell the liquid sample containing the target analyte (1) removal and magnetic elements (10) on-20 the best way to remove unbonded components a small amount of washing solution from the analysis cell (1) ge- being irritated, working electrode (5), counter electrode (6) and reference Using electrode (7), the target analyte is injected into the receptor-25 after connecting to magnetic elements (10) a second electrochemical impedance spectroscopy measurement the realization of the problem and Reference electrochemical impedance spectroscopy measurement. with second electrochemical impedance spectroscopy öl-30 Based on the difference between the two, the target analyte exists. Determining the extent and / or quantity. 31 30. A method according to claim 29, containing the target analyte. before the liquid sample is transferred to the analysis cell (1); Protein and fat originating from the food matrix found in the sample, pigments, particles and / or other interfering components To reduce the adsorption rate, the sample must be 5% adsorbed and / or separator. from a chemical purification section containing material- A method characterized by its use.

31. Method according to claim 29 or 30, target analyte before the liquid sample containing is transferred to the analysis cell (1) first, 10 multiple liquid packages with specific volumes separation and magnetic element in each liquid packet- by passing through the region where (10) is located separately The method being characterized.

32. The method is in accordance with claim 31, and each liquid package inside the analysis cell (1), with the liquid inlet section (3) and 15 a return circulation line between the liquid outlet section (2) through or by applying positive and negative pressure repeated back-and-forth movement a method characterized by its...

33. The method is according to any of claims 29 to 32, 20 magnetic elements of the liquid sample containing the target analyte (10) during the passage through the region where it is located, analysis flow narrowing of the flow cross-section inside cell (1) local reduction with the wall (8) and thus liquid The sample was oriented via magnetic elements (10)-25 a method characterized by its application.

34. The method is according to any of claims 29 to 33, gas bubbles inside the analysis cell (1) from the working electrode (5), from the counter electrode (6) and a 30 positioned away from the reference electrode (7) by directing the bubble collecting element (9) character- The method described. 32 35. The method is according to any of claims 29 to 34, target analyte mycotoxin, aflatoxin, ochratoxin, de- oxynivalenol, zearalenone, fumonisin, patulin, pesticide, antibiotic residue, veterinary drug residue, allergens gene, protein, microorganism, DNA, RNA or small mo-5 A method characterized by the presence of at least one lecule.

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