TWM462368U - Electrochemical biosensor without hematocrit correction - Google Patents

Abstract

This invention is related to an electrochemical biosensor free from the influence of hematocrit effect on blood measurement. The electrochemical biosensor comprises a special core-shell conductive polymer silk-liked structure, which was porous and immobilized in wave-formed micro fluidic channel of electrochemical biosensor. The pressure sensitive core-shell polymer was conductive and a non-contact induced electrical field was generated in micro fluidic channel while the specific voltage being applied to the polymer. Before the blood flowing into the reaction zone of the biosensor, the charged particles, such as red blood cell or other viscous matters in blood, will be trapped and screened in micro fluidic channel. The pretreatment will prevent the deviation of measurement from hematocrit effect.

Description

Electrochemical biosensor without correction of blood volume ratio effect

This creation is related to an electrochemical biosensor that is free of the Hematocrit effect (Hct), especially when the electrochemical biosensor measures the concentration of the analyte in the blood. Non-contact electric field induction method, combined with a special internal conductive core-shell pressure-sensitive polymer winding wire-like conductive structure in a curved micro-channel structure, pre-charged red blood cells or impurities in the blood Excavating and screening in the curved micro-fluid structure of the electrochemical biosensor, and filtering out some components that cause the blood viscosity to rise, and avoiding entering the contact reaction electrode region of the electrochemical biosensor, In order to achieve the purpose of eliminating the interference of blood volume ratio effect.

The use of electrochemical biosensors is quite convenient for various blood indicators of patients, such as blood sugar, uric acid, cholesterol, and the like. At present, in addition to the repetitive use of sensors commonly used in medical institutions or inspection agencies, disposable disposable electrochemical sensors for commercial applications are also popular with non-professional medical personnel or home care users.

The structure of the conventional electrochemical biosensor is mostly composed of a simple printed electrode and a multi-layer substrate having a linear microchannel and an electrochemical bioreactor region, and the measured value of the body fluid can be quickly and conveniently obtained. Mainly blood, then In practical applications, the detection of small amounts of blood is still limited by the microscopic flow resistance of a small amount of finite reaction material (less than 3 μl, μL) in the local reaction interval, which leads to errors in the measurement results. Usually the normal normal blood volume ratio of adults is about 30%-50%, but from the actual measurement statistics, there is also a blood volume ratio between 20-75%, like the blood ratio of the average baby. That is between 50% and 75%.

From the experimental analysis results, the higher blood volume ratio results in too low blood analyte measurement results, whereas the lower blood volume ratio makes the measurement value higher. The main reason is that when the blood volume ratio is high, the red blood cells will hinder the chemical reaction on the electrochemical biosensor, resulting in a low current response, and a low blood volume ratio, and vice versa, the current response will be high. Since the change of blood volume ratio is common in different blood, which affects the accuracy of measurement, the error caused by the blood volume ratio effect on the electrochemical biosensor is overcome, and the blood of different blood volume ratio conditions is timely Compensation correction has become a goal of the development of most commercial electrochemical biosensors, and an electrochemical sensing system with blood volume compensation function has been established.

A prior art practice, such as U.S. Patent No. 7,407,811 ('811), discloses a method of reducing blood volume ratio interference when measuring the concentration of an analyte in blood, wherein the method of measuring the blood volume ratio is to apply an alternating signal to the measurement. In the blood in the electrochemical biosensor, and measure the phase angle and the admittance magnitude of the reaction when the AC signal responds, and then match the formula Import and detect the blood volume ratio. The '811 patent, by its method of measuring the blood volume ratio, further proposes a blood analyte to be measured method for correcting the blood volume ratio function. However, the method of adjusting the blood volume ratio of the blood measurement value disclosed in the '811 patent provides an alternating current and a direct current signal to the body fluid of the reaction zone of the electrochemical biosensor, and uses a contact reactive electrode set for detection. In addition, because the DC signal and the AC signal are in the same reaction zone and the same reaction electrode group, it is easy to generate noise interference or mutual interference, and the temperature correction is needed to obtain the correct blood sugar value, and the direct application on the reaction electrode. The specific current method also causes some of the electrochemically active substances in the blood to participate in the reaction, thereby disturbing the actual measurement results. In addition, the above-mentioned exposure to the application of alternating current and direct current on the reaction electrode prolongs the electrochemical reaction time of the blood. And to increase the complexity of the measurement steps, summed up the above problems, such as the method disclosed in '881 is still the need for improvement.

In order to overcome the effect of the blood volume ratio effect on the electrochemical biosensor to measure the analyte in the blood, and to improve the lack of the above conventional correction compensation method, the present invention provides a method that does not require direct application of current in the creation. The non-contact electric field senses the surface of the electrode layer and constitutes an electrochemical biosensor structure with an uncorrected blood volume ratio, which is designed by using a curved micro-channel structure and utilizing an internal conductive core-shell type Pressure-sensitive polymer winding wire-like conductive structure into filling and filling Set in a curved microfluidic structure, when a DC or AC voltage signal is applied, a non-contact induced electric field is formed in the curved microfluidic structure, and the blood enters the electrochemical biosensing. In the curved micro-flow channel structure, the charged substance in the blood can be screened and filtered, and some components that cause the blood viscosity to rise are filtered out, thereby avoiding the blood volume ratio effect on the electrochemical sensor. The impact of the measurement, thereby improving the accuracy of the electrochemical biosensor.

In order to achieve the above-mentioned effect of overcoming the blood volume ratio effect on the measurement of the analyte in the blood, and obtaining an accurate measurement result of the analyte in the blood, one of the technical methods of the present invention is to provide an uncorrected blood volume ratio effect. An electrochemical biosensor for measuring a concentration of an analyte in a blood, comprising: a substrate having a curved microfluidic structure for use as a blood into an electrochemical biosensor; A conductive layer structure is mainly composed of a set of non-contact electric field sensing electrode layers and another set of contact reactive electrode layers, and the non-contact electric field sensing electrode layers are distributed on both sides of the curved microfluidic structure. And partially electrically connected to the inner electrically conductive core-shell type pressure-sensitive polymer-wound wire-shaped conductive structure, and when a specific voltage signal is applied to the direct current or the alternating current, the curved shape of the electrochemical biosensor can be formed. In the flow channel structure, a porous charged channel in the form of an electric field is generated, and the redness of the charge can be obtained when the blood passes through the curved microfluidic structure by the siphon principle. The possible charged particles or viscous substances in the ball and blood are hindered in the porous structure in the curved micro-channel structure or in the curved surface of the curved-shaped micro-channel structure, only the blood to be analyzed and the rest body The liquid flows through the microchannel and goes to another set of contact reaction electrode layers, and the set of contact reaction electrode layers is used for detecting the concentration of the analyte in the blood; an internal conductive core-shell pressure sensitive polymer The wire-shaped conductive layer structure is arranged to cover the curved micro-channel and partially overlap with the non-contact electric field-sensing electrode layer structure, and then press the overlapping regions by mechanical pressure. Further, a pressure-sensitive conductive contact region is formed to form a contact conductive state; an insulating layer structure is accompanied by a double-sided adhesive layer for covering and fixing on the conductive layer and exposing a portion of the conductive layer to expose the conductive a layer region serving as a contact reaction electrode layer and a connector end for transmitting a voltage and current signal; an electrochemical reaction layer covering a partially uncovered contact reaction electrode layer fixed to the conductive layer as a The analyte in the blood reacts and generates a specific current signal; and an upper cap layer overlies the structure of the insulating layer to form a complete electrochemical Sensor micro-channel structure material, and only the exposed portion of the conductive layer to the connector is transmitted as the signal.

In another specific embodiment of the present invention, the conductive layer serves as a contact reaction electrode layer of the electrochemical biosensor reaction zone, and provides a working electrode and a counter electrode in addition to participating in the reaction. In addition, a trigger sensing electrode layer of a contact reaction electrode layer may be further included, and the trigger sensing electrode layer is disposed at the end of the curved micro flow channel structure to confirm whether the blood reaches and Filling the reaction zone covering the contact reaction electrode layer of the electrochemical reaction layer, and starting the electrochemical growth after the induction confirmation The working voltage signal required by the object sensor, in addition, the triggering sensing electrode layer may be connected in parallel with the working electrode layer end of the original contact reactive electrode layer after the starting working voltage signal, thereby increasing the contact reactive electrode layer. The working electrode area increases the current signal strength. Also. The non-contact electric field sensing electrode layer of the conductive layer is applied to the non-contact electric field sensing electrode layer of the conductive layer by a direct current or alternating current bias voltage signal, and is not covered by the insulating layer structure, the double-sided adhesive layer and the upper cover layer. a partial region, which is composed of a connector notch structure of an insulating layer, a connector notch structure of the double-sided adhesive layer and a connector notch structure of the upper cap layer, and can be connected with the signal transmission connection of the contact reactive electrode layer The terminals together form a set of signal connector end connection buses as an interface for applying a voltage signal to the electrochemical biosensor or collecting a voltage and current signal generated by the electrochemical biosensor; another better The method is characterized in that the contact end of the signal transmission connector of the non-contact electric field sensing electrode layer is disposed on the two sides of the electrochemical biosensor near the curved micro flow channel structure, thereby reducing the voltage signal applied by the conductive layer. Impedance loss and interference with other voltage and current signals can also be used in electrochemical bioreactors for electrochemical biosensors The signal detection device avoids the interference of high and low voltage transmission and distribution and reduces the circuit complexity in design and manufacture.

In another example of the present creation, in the curved micro-channel structure design of the electrochemical biosensor described in the present invention, the inner conductive outer core-shell pressure-sensitive polymer is wound into a filament shape. The conductive structure has a low dielectric constant with low dielectric properties The molecularly wound filament structure is substituted, and the non-contact electric field sensing electrode layer of the conductive layer is directly extended to the runner surface of the curved microfluidic structure, but the non-contact electric field sensing electrodes of the conductive layer are both The non-conductive polymer with low dielectric properties is wrapped around the filament structure and cannot be in direct electrical contact with the blood flowing through the curved microchannel structure, but can be applied with a specific AC or DC voltage signal. The non-contact electric field sensing electrode layer causes the non-contact electric field sensing electrode layer to generate a corresponding electric field in the curved microfluidic structure, so that the specific charged red blood cells or substances in the passing blood The electric fields are mutually inductive, thereby blocking the substances related to the blood volume ratio effect from being trapped in the curved microfluidic structure.

Through the above-mentioned technical means and design, when the concentration of the analyte in the blood is measured by the electrochemical biosensor proposed by the present invention, it is better to obtain the substance by causing the blood volume ratio effect. The measurement results are analyzed, and the technical means and design proposed by the present invention can also reduce the design and manufacturing complexity of the electrochemical bioreaction signal detecting device and eliminate the step of correcting the blood volume ratio, thereby effectively improving the blood to be analyzed. Convenience and timeliness of things.

This creation is related to an electrochemical biosensor that corrects the blood volume ratio effect. The electrochemical biosensor is likely to affect the blood before it is measured by electrochemical reaction in the blood. Analyte concentration measurement The correlation factor of the fruit is pretreated, such as the blood volume ratio effect in the blood, thereby reducing the deviation or error of the measurement result. The pretreatment performed in the curved microfluidic structure of the electrochemical biosensor through the present invention can exclude the blood volume ratio effect on the concentration of the analyte in the blood, such as blood glucose concentration, uric acid concentration or cholesterol value, etc. The influence of the measurement results can effectively monitor the blood health status of the tested patients for a long time, thereby avoiding the complications of chronic diseases. In order to obtain an effective and accurate measurement result, please refer to the first figure, which is shown in the designated representative figure presented for this creation. This creation provides an electrochemical biosensor without correction of the blood volume ratio effect. The invention comprises a non-contact electric field sensing electrode layer (20) and an inner conductive core-shell type pressure sensitive polymer winding wire-shaped conductive structure (30), which is fixed to the electrochemical biosensor and has a curved shape micro The substrate (10) of the flow channel structure is provided on one end of the material, and a working area of the contact reaction electrode layer (21, 22, 23) is disposed at the end of the curved microfluidic structure (11), thereby passing the curved shape. The blood of the microchannel structure (11) is pretreated, and an induced electric field effect is generated in the curved microfluidic structure (11) by applying a voltage signal, but the conduction behavior of the actual electron transfer exchange mode is not performed. In order to exclude the charged components that may produce a hematocrit effect, such as red blood cells or charged or viscous impurities in the blood, and then the blood entering the contact reaction electrode layer (21, 22, 23) working area has Unaffected by the blood volume ratio effect Sound, so that the accurate measurement results of the analyte in the blood can be obtained by the controlled electrochemical biological reaction, and the curved micro-flow channel structure (11) can also carry the particles or impurity components in the blood. Block or Blocking the curved surface structure, avoiding it flowing to the working area of the contact reaction electrode layer (21, 22, 23) at the end of the curved microfluidic structure (11), thereby interfering with the blood measurement result, and including On the conductive layer such as the contact electric field sensing electrode layer (20) and the contact reaction electrode layer (21, 22, 23), a micro flow channel structure (41) having a double-sided adhesive layer and a double-sided adhesive layer are added. The double-sided adhesive layer (40) of the connector notch structure (42) is stacked and fixed by the micro-channel structure (51) having the insulating layer and the insulating layer (50) of the connector notch structure (52) of the insulating layer, and The mechanically-pressurized core-shell-type pressure-sensitive polymer-wound filament-like conductive structure (30) is pressed against a region where the non-contact electric field sensing electrode layer (20) and the partial double-sided layer (40) overlap each other. And a non-contact electric field sensing electrode layer capable of conducting the non-contact electric field sensing electrode layer (20) and the inner conductive core-shell type pressure sensitive polymer winding wire-like conductive structure (30) and internal conductive The core-shell pressure-sensitive polymer is wound around the filament-like conductive structure and overlaps the pressure-sensitive conductive back region (511 ), but otherwise, the inner conductive core-shell pressure-sensitive polymer is wound around the filament-like conductive structure (30) without any mechanical stress, for example, into the curved micro-channel structure (11). The wound filament structure portion is completely in the state of internal conduction but external insulation, and the triggering induction of the working electrode layer (21) and the contact reaction electrode layer of the partially exposed contact reaction electrode layer above the insulating layer (40) The electrode layer (22) and the surface of the auxiliary electrode layer (23) of the contact reaction electrode layer are fixed to the electrochemical reaction layer (60), and the connector gap structure (72) having the vent hole (71) and the upper cap layer is provided. The upper cap layer (70) is overlaid thereon to form a complete and functional, electrochemical electrochemical sense as shown in the third figure Schematic diagram of the combination of detectors.

The term "voltage signal" is defined as the result of superimposing or superimposing a separate signal covering an alternating or direct current potential or an alternating current and direct current potential signal; the signal may also be a superposition or superposition of a pulsed signal plus a direct current potential.

In the first implementation case of this creation, an electrochemical biosensor is provided which is free of the corrected blood volume ratio effect. Please refer to the first and third figures, which is the electromagnetization effect of the artificial blood volume ratio effect. A perspective exploded view and a combined schematic view of an embodiment of a student sense sensor, comprising a substrate (10) having a curved microfluidic structure, a curved microfluidic structure (11), and a non-contact Electric field induction electrode layer (20), working electrode layer (21) of contact reaction electrode layer, trigger induction electrode layer (22) of contact reaction electrode layer, auxiliary electrode layer (23) of contact reaction electrode layer, internal Conductive core-shell type pressure-sensitive polymer-wound wire-like conductive structure (30), double-sided adhesive layer (40), micro-channel structure of double-sided adhesive layer (41), connector gap structure of double-sided adhesive layer ( 42), insulating layer (50), microchannel structure (51) of insulating layer, connector notch structure (52) of insulating layer, electrochemical reaction layer (60), upper cap layer (70), vent hole (71) And a connector gap structure (72) of the upper cover layer.

The substrate (10) having a curved micro flow channel structure may be a non-conductive material, and a curved deep concave structural groove is formed on the surface thereof for use as a micro flow channel.

The non-contact electric field sensing electrode layer (20), the working electrode layer (21) of the contact reactive electrode layer, the trigger sensing electrode layer (22) of the contact reactive electrode layer, and the auxiliary electrode layer of the contact reactive electrode layer (23) Directly covering a substrate (10) having a curved microfluidic structure and having a curved microfluidic structure (11) One side, and distributed on the same plane, wherein the non-contact electric field sensing electrode layer (20) is close to the introduction side of the blood to be tested, and the remaining electrode layers are with one end and the electrochemical reaction layer (60) The other end serves as a contact connection electrochemical bioreactor signal detecting device; and the triggering sensing electrode layer (22) of the contact reactive electrode layer designed by the present invention is not only used as an application for confirming whether the blood to be analyzed is sufficient. It can be used as the working electrode of the second group of electrochemical reactions in the subsequent electrochemical bioreaction, and simultaneously provide the same electrochemical reaction voltage signal with the working electrode layer (21) of the contact reaction electrode layer to enhance the electrification. The sensitivity of the student sensor detection.

The inner conductive core-shell pressure-sensitive polymer-wound wire-shaped conductive structure (30) is fixed in the curved shape micro-channel structure (11) by a squeezing filling method and partially and non-contact electric field sensing electrode layer (20) One side of the adjacent curved micro-channel structure (11) partially overlaps, as shown in the second figure, the overlapping portion becomes the non-contact electric field sensing electrode layer and the interior as shown in FIG. The conductive core-shell type pressure-sensitive polymer is wound around the filament-like conductive structure and the pressure-sensitive conductive adhesion region (511), and the inner conductive core-shell pressure-sensitive polymer-wound filament-shaped conductive structure (30) has a curved shape. The length of the intrusion on the substrate (10) of the microchannel structure is mainly the working electrode layer (21) which does not cover the contact reaction electrode layer.

In combination with an electrochemical biosensor that corrects the blood volume ratio effect proposed in the first embodiment of the present invention, the non-contact electric field sensing electrode layer (20) is internally conductive and core-shell pressure sensitive polymer Winding wire-shaped conductive structure (30) partially heavy After the overlay is covered, it is partially covered with a double-sided adhesive layer (40), and then covered with an insulating layer (50) of the same shape area to be fixed and further subjected to internal pressure-sensitive core-shell pressure by mechanical pressure. The polymer-wound filament-like conductive structure (30) is pressed together with a non-contact electric field sensing electrode layer (20) and a portion of the double-sided back layer (40), but is electrically conductive in the interior of the plug-in filling. The curved type microfluidic structure (11) of the pressure-sensitive polymer-wound filament-like conductive structure (30) is in an insulating state, and can only be applied to the non-contact electric field sensing electrode layer by applying a voltage signal (20). An induced electric field is generated between the surface of the curved microchannel structure (11) and the insulating outer layer of the internally conductive core-shell pressure-sensitive polymer-wound filament-like conductive structure (30). A schematic side view of the inlet end of the microchannel is shown in the third figure. As can be seen from the figure, the blood to be detected can only pass through the core of the curved microfluidic structure (11) which is not electrically conductive. The space occupied by the pressure sensitive polymer wrapped wire-like conductive structure (30), As can be seen from the fifth figure, when blood flows through the curved microfluidic structure (11), the direction of the flow path changes, and the charged or heavier, more viscous components such as red blood cells may be retained in the blood. In the microchannel curvature, this design enhances the blocking of blood impurities or red blood cell components of different sizes that may affect the blood volume ratio effect.

In the actual measurement of this creation, when measuring the analyte in the blood, taking the blood glucose concentration as an example, before introducing the blood into the curved micro-channel structure (11) in the electrochemical biosensor, Provides a constant current or AC superposition voltage signal applied to the non-connected The contact electric field sensing electrode layer (20) causes an inner electrically conductive core-shell type pressure sensitive polymer winding wire-like conductive structure (30) in the curved micro flow channel structure (11) to generate an inductive electric field as a attracting And adsorbing a specific charged component in the blood, and the curved microchannel structure (11) may include, but is not limited to, a curved form of a two-dimensional plane or a depth of the curved micro-flow structure (11) Some have three-dimensional curved structures formed by different high and low wave patterns. When the blood that has been flowed through the curved microfluidic structure (11) and is pretreated by the induced electric field passes through the internally conductive core-shell type pressure sensitive polymer wrapped around the filamentary conductive structure (30), the red blood cells and blood are excluded. The charged or viscous particulate impurities, which may affect the blood volume ratio effect, can be further transported to the working electrode layer (21) of the contact reaction electrode layer, the triggering electrode layer (22) of the contact reaction electrode layer, and the contact The electrochemical bioreactor region composed of the auxiliary electrode layer (23) of the reaction electrode layer is as shown in Fig. 5, and is covered by the electrochemical reaction layer (60).

When the pre-treated blood reaches the triggering electrode layer (22) of the contact reaction electrode layer as shown in FIG. 5, the auxiliary electrode layer (23) and the contact reaction electrode of the contact reaction electrode layer are transmitted through the blood. The conductive path between the triggering sensing electrode layer (22) of the layer is formed, so that the electrochemical electrochemical reaction signal detecting device is fed back by the voltage signal generated by the conductive path, and the working electrode layer of the contact reactive electrode layer (21) The triggering electrode layer (22) of the contact reaction electrode layer and the auxiliary electrode layer (23) of the contact reaction electrode layer extend to the signal connector end contact pin at one end of the electrochemical biosensor, as shown in the third figure. As shown, away from bending The end face of the microfluidic channel (11) sends the detection signal back to the electrochemical bioreactor signal detecting device.

In the region covered by the electrochemical reaction layer (60), the detection signal of the triggering sensing electrode layer (22) of the contact reaction electrode layer is used to confirm that a sufficient and accurate blood sample amount to be analyzed has been introduced, and then immediately The working electrode layer (21) of the contact reaction electrode layer, the triggering electrode layer (22) of the contact reaction electrode layer and the auxiliary electrode layer of the contact reaction electrode layer are provided by the combined electrochemical bioreactor signal detecting device (23) a signal connector end contact extending to one end of the electrochemical biosensor, as shown in the third figure, required to apply an electrochemical bioreaction away from the end face of the curved microchannel (11) The working voltage, wherein the working electrode layer (21) of the contact reactive electrode layer and the triggering sensing electrode layer (22) of the contact reactive electrode layer are connected in parallel to increase the electrode area of the working electrode layer to enhance the electrochemical reaction layer (60) Signal sensitivity of the reaction zone.

In the second implementation case of this creation, an electrochemical biosensor is provided which is free of the corrected blood volume ratio effect. Please refer to the sixth and eighth figures, which is an electrochemical method for correcting the blood volume ratio effect. An exploded perspective view and a combined schematic view of an embodiment of a biosensor, comprising a substrate (10a), a non-contact electric field sensing electrode layer (20a), a working electrode layer (21a) of a contact reactive electrode layer, and a contact The trigger electrode layer (22a) of the reaction electrode layer, the auxiliary electrode layer (23) of the contact reaction electrode layer, the double-sided adhesive layer (30a), the micro flow channel structure (31a) of the double-sided adhesive layer, and the double-sided layer Connector gap structure (32a), insulating layer (40a), insulating layer Microchannel structure (41a), connector gap structure (42a) of insulating layer, non-conductive polymer-wound filament structure (50a), electrochemical reaction layer (60a), upper cap layer (70a), vent hole (71a) And a connector gap structure (72a) of the upper cover layer.

In the second embodiment of the present invention, on the same plane above the substrate (10a), the non-contact electric field sensing electrode layer (20a), the working electrode layer (21a) of the contact reactive electrode layer, The triggering electrode layer (22a) of the contact reaction electrode layer and the auxiliary electrode layer (23a) of the contact reaction electrode layer are directly covered, wherein the non-contact electric field sensing electrode layer (20a) is close to the blood test body fluid to be tested. The introduction side, while the other electrode layer is in contact with the electrochemical reaction layer (60a) at one end thereof, and the other end is used as a contact connection electrochemical bioreaction signal detecting device; and the contact reaction separately designed by the present invention The triggering electrode layer (22a) of the electrode layer is not only used as an application for confirming whether the blood to be analyzed is sufficient, but also can be regarded as a second group of electrochemical working electrodes and working with the contact reactive electrode layer in the subsequent electrochemical biological reaction. The electrode layer (21a) simultaneously provides the same electrochemical reaction voltage signal to enhance the detection sensitivity of the electrochemical biosensor.

Further, in conjunction with the exploded assembly diagram of the sixth figure, the double-sided adhesive layer (30a) can be partially covered on the substrate (10a) having the conductive layer covering configuration, and only the micro flow channel structure of the double-sided adhesive layer is exposed ( 31a) and a portion of the connector gap structure (32a) of the double-sided adhesive layer, and then the insulating layer (40a) is subsequently fixed on the double-sided adhesive layer (30a) in the same covering shape, and will also be fixed thereon Insulating layer The microchannel structure (41a) and the connector gap structure (42a) of the insulating layer are exposed and are not completely covered, and are used to form a curved microfluid used as a blood introduction by the present electrochemical biosensor. The contact area between the channel structure and the electrochemical reaction, and the contact pin of the signal connector end, thereby connecting to the electrochemical bioreactor signal detecting device and transmitting the voltage signal.

The adhesive layer (30a) and the insulating layer (40a) are partially covered on the non-contact electric field sensing electrode layer (20a), and only a part of the design is exposed as a contact portion of the connector interface, as shown in FIG. 7 One end of the curved microfluidic structure is covered to make it in an insulating condition, but its edge remains as a curved microfluidic structure. As shown in the seventh figure, the non-contact electric field sensing electrode layer (20a) is coated by the double-sided adhesive layer (30a) and is not in contact with the non-conductive polymer-wound filament structure (50a) in the curved microchannel, and when a voltage signal is applied to the non-contact electric field When the electrode layer (20a) is sensed, only an induced electric field is generated near the wall surface of the micro flow channel, and the induced electric field will not extend to the non-conductive polymer winding filament structure (50a), such as the micro flow channel of the ninth figure. A schematic side cross-sectional view of the entrance end of the structure, the non-contact electric field sensing electrode layer (20a) is covered by the double-sided adhesive layer (30a) and the insulating layer (40a) to form an insulated microfluidic structure.

In the second practical case of the present invention, in the curved microfluidic structure formed by stacking the adhesive layer (30a) and the insulating layer (40a), the filament structure (50a) is wound with a non-conductive polymer. And the electrochemical reaction layer (60a) is covered and fixed in different regions of the microchannel, wherein the non-conductive polymer-wound filament structure (50a) is placed close to the non- In the microchannel region of the contact electric field sensing electrode layer (20a), when the blood to be analyzed is introduced into the microchannel, the charged red blood cells or the viscous particles in the blood can be carried out by the induced electric field applied near the microchannel. Inducing attraction and accumulating on the curved surface of the microchannel, as shown in the microchannel structure of the tenth diagram and the schematic view of the electrode layer, and in addition, the non-conductive polymer winding filament structure (50a) in the microchannel The remaining red blood cells or particulate impurities that are not attracted by the electric field can be filtered to prevent them from entering the micro-fluid channel as a region of the fixed electrochemical reaction layer (60a), thereby generating a blood volume ratio effect.

The electrochemical reaction layer (60a) is fixed and covers the working electrode layer (21a) of the contact reaction electrode layer, the trigger sensing electrode layer (22a) of the contact reaction electrode layer, and the auxiliary electrode layer of the contact reaction electrode layer ( 23) A region in the microchannel that is not covered by the double-sided adhesive layer (30a) and the insulating layer (40a) as an end point when blood is introduced into the microchannel and can transmit an electrochemical reaction signal from the electrode to the electrochemical Bioreactive signal detection device.

The upper cap layer (70a) covers the insulating layer (40a) and includes a vent hole (71a), and the vent hole (71a) is disposed at the other end of the blood to be analyzed corresponding to the curved microfluidic structure. That is, in the microchannel, above the triggering electrode layer (22a) of the contact reaction electrode layer covered by the bonding layer (30a) and the insulating layer (40a), the microfluid can be quickly introduced when the blood is introduced into the microchannel. The air in the channel is discharged, shortening the measurement time of the electrochemical biosensor. The upper cap layer (70a) also has a connector notch structure (72a) formed on the upper cap layer (70a) for introducing the upper cap layer of the blood end to be analyzed. The structure cooperates with the exposed portion of the non-contact electric field sensing electrode layer (20a) for applying a voltage signal and generating an induced electric field around the curved microchannel.

In summary, the main advantages and characteristics of an electrochemical biosensor without the blood volume ratio effect proposed by the present invention can be seen in the following specific test results and patent application scope.

Specific test results

The following specific implementation test results are used to demonstrate the present invention. The test results of this specific implementation are not intended to limit the scope of the present invention in any way or concept, but are used to indicate how to implement the material and method of the present invention.

The test of the specific implementation adopts the electrochemical biosensor structure of the uncorrected blood volume ratio effect proposed by the present invention, and the specific electrochemical biosensor structure of the first embodiment case and the second embodiment case are treated. The blood glucose concentration in the blood is detected, and the concentration measured by the YSI is compared, and the DC voltage signal (25V) is applied as the electrochemical biosensor measurement result after the non-contact inductive electric field. During the test, blood samples to be analyzed with different blood volume ratio conditions (HCT, %) were prepared, and the actual measurement of blood glucose concentration was performed as an electrochemical biosensor that provided YSI and the artificial blood volume ratio-free effect of the creation. According to the results of the measurement, the difference between the electrochemical biosensor and the YSI measurement blood glucose value of the artificial blood-free ratio effect of the creation is compared by comparing the applied voltage or not, and the test methods are compared. CV (coefficient of variation, Coefficient of variation) and deviation from YSI (dev.of YSI), and the test results are shown in Table 1 and Table 2, wherein Table 1 is the electrochemical biosensor structure using the first embodiment described above. The difference between the CV (deviation coefficient) and the blood glucose concentration and the YSI measurement of the blood glucose concentration value under different blood volume ratio conditions is measured, and the electrochemical biosensor structure of the second embodiment is used in the second embodiment. The relative result of the measurement. It can be seen from the results of the table that, regardless of the electrochemical biosensor of the first or second embodiment, when the voltage signal is applied as the pre-corrected blood volume ratio effect, the results are effective to improve the accuracy of measuring blood glucose. (dev.of YSI<4), it is also very obvious to eliminate the difference in blood glucose concentration measurement caused by the change of blood volume ratio of the blood to be analyzed (CV<3.0), and it can achieve no need for blood volume. Accurate and accurate actual blood glucose concentration can be obtained by deviation or compensation correction.

Different modifications and variations of the method or structure of the present invention will obviously not depart from the scope and spirit of the present invention. Although the creation has described specific better specific facts or implementation cases, it must be understood that the creation should not be unduly restricted to such specific facts or implementation cases. In fact, the various modifications, structures and methods of the present invention, which are obvious to those skilled in the art, are also covered by the following claims.

(10) ‧‧‧Substrate with curved microfluidic structure

(11)‧‧‧Bend type microchannel structure

(20) ‧‧‧ Non-contact electric field sensing electrode layer

(21) ‧‧‧Working electrode layer of contact reaction electrode layer

(22) ‧‧‧Tactile sensing electrode layer of contact reaction electrode layer

(23) ‧‧‧Auxiliary electrode layer of contact reaction electrode layer

(30)‧‧‧Internal conductive core-shell pressure-sensitive polymer wrapped filamentary conductive structure

(40) ‧ ‧ double-sided layer

(41) ‧‧‧Microfluidic structure of double-sided layer

(42) ‧‧‧ Connector gap structure of double-sided layer

(50)‧‧‧Insulation

(51) ‧‧‧Microfluidic structure of the insulating layer

(511) ‧ ‧ 感 电场 电场 电场 电场 非 非 ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧ ‧

(52) ‧‧‧ Connector gap structure

(60) ‧‧‧Electrochemical reaction layer

(70) ‧ ‧ upper cover

(71) ‧ ‧ vents

(72) ‧‧‧Connector gap structure

(10a)‧‧‧ Non-conductive substrate

(20a) ‧‧‧ Non-contact electric field sensing electrode layer

(21a) ‧‧‧Working electrode layer of contact reaction electrode layer

(22a) ‧‧‧Tactile sensing electrode layer of contact reaction electrode layer

(23a) ‧‧‧Auxiliary electrode layer of the contact reaction electrode layer

(30a) ‧ ‧ double-sided layer

(31a) ‧‧‧Microfluidic structure of double-sided layer

(32a)‧‧‧ Connector gap structure of double-sided layer

(40a)‧‧‧Insulation

(41a) ‧‧‧Microfluidic structure of the insulating layer

(42a) ‧‧‧Insulator connector gap structure

(50a)‧‧‧ Non-conductive polymer winding filament structure

(60a) ‧‧‧Electrochemical reaction layer

(70a) ‧ ‧ upper cover

(71a) ‧ ‧ vents

(72a) ‧‧‧Connector gap structure

The first figure is an exploded perspective view of a first embodiment of the present electrochemical biosensor.

The second figure is a schematic diagram of the electrode layer of the first figure of the present invention and the inner core of the core-shell pressure-sensitive polymer-wound wire-shaped conductive structure.

The third figure is a combined schematic diagram of the first embodiment of the present electrochemical biosensor.

The fourth figure is a side cross-sectional view of the inlet end of the microchannel of the third figure of the creation.

The fifth figure is a schematic cross-sectional view of the micro-channel structure of the third figure of the present creation.

The sixth figure is a perspective exploded view of a second embodiment of the present electrochemical biosensor.

The seventh figure is a schematic diagram of the assembly of the electrode layer of the sixth figure and the non-conductive polymer winding wire structure.

The eighth figure is a combined schematic diagram of the second embodiment of the present electrochemical biosensor.

The ninth figure is a side cross-sectional view of the inlet end of the microchannel of the eighth drawing of the present creation.

The tenth figure is a schematic view of the microchannel structure and the upper side of the electrode layer in the eighth drawing of the present creation.

(10) ‧‧‧Substrate with curved microfluidic structure

(11)‧‧‧Bend type microchannel structure

(20) ‧‧‧ Non-contact electric field sensing electrode layer

(21) ‧‧‧Working electrode layer of contact reaction electrode layer

(22) ‧‧‧Tactile sensing electrode layer of contact reaction electrode layer

(23) ‧‧‧Auxiliary electrode layer of contact reaction electrode layer

(30)‧‧‧Internal conductive core-shell pressure-sensitive polymer wrapped filamentary conductive structure

(40) ‧ ‧ double-sided layer

(41) ‧‧‧Microfluidic structure of double-sided layer

(42) ‧‧‧ Connector gap structure of double-sided layer

(50)‧‧‧Insulation

(51) ‧‧‧Microfluidic structure of the insulating layer

(52) ‧‧‧ Connector gap structure

(60) ‧‧‧Electrochemical reaction layer

(70) ‧ ‧ upper cover

(71) ‧ ‧ vents

(72) ‧‧‧Connector gap structure

Claims (10)

An electrochemical biosensor for correcting blood volume ratio effect, which is used for measuring the concentration of an analyte to be tested in blood, comprising: - a substrate having a curved shape microchannel structure; - a conductive layer, Covering the substrate and comprising a non-contact electric field sensing electrode layer and a working electrode layer of the contact reactive electrode layer, a triggering sensing electrode layer of the contact reactive electrode layer and an auxiliary electrode layer of the contact reactive electrode layer, wherein The non-contact electric field sensing electrode layer is mainly used to introduce blood into the electrochemical biosensor to eliminate the influence of the blood volume ratio effect on the accuracy of the measurement result, and other contact reaction electrode layers are used to measure the blood. The concentration of the analyte to be analyzed; - an internally conductive core-shell type pressure-sensitive polymer-wound filament-like conductive structure, which is filled in a curved microfluidic structure and partially connected to the non-contact electric field sensing electrode layer The overlapping sensitivities then form a conductive path and form an induced electric field distribution in the curved microfluidic structure when a voltage signal is applied to the non-contact electric field sensing electrode layer. In order to block the flow of charged or viscous impurities in the red blood cells or blood; - a double-sided adhesive layer partially covering the conductive layer to cover the pressure-sensitive conductive region of the insulating curved micro-channel structure and exposed The remaining conductive layer serves as one end of the connector voltage signal transmission and as a region required for electrochemical biological reaction; - an insulating layer covering the double-sided adhesive layer, which is uniform in size, shape and size. When bonded to the double-sided adhesive layer by a mechanical pressure, the inner conductive core-shell type is also completed at the same time. The sensible pressure of the pressure-sensitive polymer-wound filament-like conductive structure and the non-contact electric field-sensing electrode layer is followed by forming a conductive path; - an electrochemical reaction layer covering the contact-reaction electrode layer portion without being double-sidedly laminated The inner region of the microchannel occupied by the core-shell pressure-sensitive polymer-wound filament-shaped conductive structure covered by the insulating layer and not electrically conductive, and in direct contact with the contact reaction electrode layer and reacting with the analyte in the blood ;- an upper cap layer covering the uppermost part of the electrochemical biosensor, which has a vent hole located at a relative position above the electrochemical bioreactor layer, and covers the curved microfluidic structure but only exposes As the conductive layer area for the signal connector end contact pin. An electrochemical biosensor for correcting blood volume ratio effect according to claim 1, wherein the substrate having the curved microfluidic structure can be printed, coated, and transferred. Physical or chemical means such as etching, overmolding, mechanical cutting or laser laser melting, further forming a structure in the form of a curved groove on an insulating substrate or a composite substrate having no surface conduction. An electrochemical biosensor for correcting blood volume ratio effect according to claim 1, wherein the conductive layer can be divided into a non-contact electric field sensing electrode layer that does not provide electron exchange transfer and provide electronic exchange transfer. Contact reaction An electrode layer including a working electrode layer, a trigger sensing electrode layer, an auxiliary electrode layer, and the like. An electrochemical biosensor for correcting blood volume ratio effect according to claim 3, wherein the electrode layer shape may be exposed as a non-contact electric field sensing electrode layer not providing electron exchange transfer Part of the region is on both sides of the curved microchannel of the electrochemical biosensor, and the voltage signal can be directly applied directly to the microchannel without extending the non-contact electric field sensing electrode layer to the electrochemical biosensor. Keep away from one end of the blood inlet. An electrochemical biosensor for correcting blood volume ratio effect according to claim 1, wherein the inner conductive core-shell pressure sensitive polymer is wound with a filament-like conductive structure, which is two layers high inside and outside. a entangled core-shell structure composed of molecules, the inner layer is mainly composed of a metal, a metal-polymer mixture, a conductive polymer or a polymer mixture containing conductive carbon, and the like, and the outer layer is a pressure-containing material. Creoles, polyalcohols, or composites with pressure-sensitive conductivity and subsequent properties. An electrochemical biosensor for correcting blood volume ratio effect, which is used for measuring the concentration of an analyte to be tested in blood, comprising: - a substrate; - a conductive layer covering the substrate and including a Non-contact electric field sensing electrode layer and working electrode layer of contact reactive electrode layer, contact reactive electrode The triggering electrode layer of the layer and the auxiliary electrode layer of the contact reactive electrode layer, wherein the non-contact electric field sensing electrode layer is mainly used for introducing blood into the electrochemical biosensor, and eliminating the blood volume ratio effect to accurately measure the result The influence of sex, while other contact-reactive electrode layers are used to measure the concentration of the analyte in the blood; - a double-sided layer is partially covered on the conductive layer to form a structure having a curved microfluidic shape, and Exposing the remaining conductive layer as one end of the signal connector end contact pin and the area required for electrochemical bioreaction; - an insulating layer covering the double-sided adhesive layer, its size, shape, size Consistently, when bonding with the double-sided adhesive layer, a complete curved micro-fluid structure is formed for blood introduction and reaction, and a voltage signal is applied to the non-contact electric field sensing electrode layer. An induced electric field distribution may be formed on the wall surface of the curved microfluidic structure to block the flow of charged or viscous impurities in the red blood cells or blood; - a non-conductive polymer a wire-wound structure which is filled in a curved micro-channel structure formed by a double-sided adhesive layer and an insulating layer, and does not form a conductive path with the non-contact electric field sensing electrode layer; - an electrochemical reaction a layer covering a region of the microchannel which is not covered by the double-sided adhesive layer and the insulating layer and which is not occupied by the non-conductive polymer-wound filament structure, and the contact reaction electrode layer Direct contact and reaction with the analyte in the blood; - an upper cap layer covering the uppermost portion of the electrochemical biosensor, having a venting hole located at a relative position above the electrochemical bioreactor layer, and covering the curved microfluidic structure but exposing only The signal connector end contacts the conductive layer area for the pin. An electrochemical biosensor for correcting blood volume ratio effect according to claim 6, wherein the non-conductive polymer is wound around a filament structure, and the filament is composed of a polymer having a low dielectric constant. The wound structure is mainly a material having a fibrous porous material such as a polymer mixture, a polymer adhesive mixed porous inorganic compound, or a porous foam polymer mixture. An electrochemical biosensor for correcting the blood volume ratio effect according to claim 6, wherein the structure of the curved micro flow channel may not be filled with the non-conductive polymer winding. The filamentous structure, only the microchannel as the direct introduction of the blood to be analyzed and the electric field induced by the wall surface of the microchannel, will attract and exclude the charged red blood cells and impurities in the blood which affect the blood volume ratio effect. An electrochemical biosensor with an anti-corrected hematocrit effect as described in claim 1 or 6 of the patent application, wherein the electrochemical biosensor structure designed to cope with the blood volume ratio-free effect is designed The measurement technique is for detecting the concentration of the analyte in the blood, comprising: providing an electrochemical biosensing of an uncorrected blood volume ratio effect as described in claim 1 or 6 of the patent application. Providing the blood into the curved microchannel with contact with the infiltrated filament structure and simultaneously applying a voltage signal to the non-contact electric field sensing electrode; providing a sweep The voltage signal is applied between the working electrode layer of the contact reaction electrode layer and the triggering electrode layer of the contact reaction electrode layer, and after the blood flows into the electrochemical reaction layer and causes short circuit between the electrodes, a DC voltage signal is applied to the contact. On the working electrode layer of the reaction electrode layer and the auxiliary electrode layer of the contact reaction electrode layer, the working electrode layer of the contact reaction electrode layer and the triggering electrode layer of the contact reaction electrode layer are simultaneously connected in parallel; The reaction signal of the reaction layer is used to obtain an actual reaction value and the concentration of the analyte in the blood is calculated. An electrochemical biosensor for correcting blood volume ratio effect according to claim 1 or 6, wherein the applied voltage signal comprises a layer applied as a non-contact electric field sensing electrode to produce Inductive electric field includes but is not limited to DC, AC, pulse and voltage signals containing various types of wave lines ranging from -50V to +50V, and applied to the contact reaction electrode layer and acting on the electrochemical reaction layer - DC voltage signal in the range of 1.5V to +1.5V.