WO2005054839A1 - ヘマトクリット(Hct)の測定方法およびそれに用いるセンサならびに測定装置 - Google Patents
ヘマトクリット(Hct)の測定方法およびそれに用いるセンサならびに測定装置 Download PDFInfo
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- WO2005054839A1 WO2005054839A1 PCT/JP2004/018019 JP2004018019W WO2005054839A1 WO 2005054839 A1 WO2005054839 A1 WO 2005054839A1 JP 2004018019 W JP2004018019 W JP 2004018019W WO 2005054839 A1 WO2005054839 A1 WO 2005054839A1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/483—Physical analysis of biological material
- G01N33/487—Physical analysis of biological material of liquid biological material
- G01N33/49—Blood
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
- G01N27/3271—Amperometric enzyme electrodes for analytes in body fluids, e.g. glucose in blood
- G01N27/3274—Corrective measures, e.g. error detection, compensation for temperature or hematocrit, calibration
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/483—Physical analysis of biological material
- G01N33/487—Physical analysis of biological material of liquid biological material
Definitions
- the present invention relates to a method for measuring Hct, a sensor used therefor, and a measuring device.
- the Hct value of blood is measured as one of the indices for knowing the properties of blood (such as blood dryness and anemia). Also, in the measurement of blood components such as glucose concentration (blood sugar level) in blood, the measured value may fluctuate depending on the Hct value, so that measurement of the Hct value may be necessary for correction.
- the Hct value can be measured by, for example, sucking blood with an anticoagulant into a capillary tube, sealing one of them with a putty, etc., centrifuging at high speed, and calculating the ratio from the height of the red blood cell layer with the whole as 100% The method (micro hematocrit method) is common.
- red blood cells are regarded as electrical pulses, and the sum of these magnitudes is calculated, and a method of automatically calculating from the average volume of red blood cells and the number of red blood cells.
- the standard Hct value for adult boys is 39-50%
- the standard Hct value for adult girls is 36-45%.
- Patent Document 1 Patent No. 3369183
- the present invention provides a method for electrochemically measuring a Hct value using a sensor.
- An object of the present invention is to provide a method of measuring Hct value excellent in measurement accuracy and reliability, and a sensor and a measuring device used for the method.
- a measurement method of the present invention is an electrochemical measurement method of blood hematocrit (Hct) value, comprising preparing an electrode system having a working electrode and a counter electrode, and No redox substance was placed on the working electrode of the two electrodes, and a redox substance was placed on the counter electrode, blood was introduced into the electrode system, and a voltage was applied to the electrode system in this state.
- Hct blood hematocrit
- the senor of the present invention is a sensor for electrochemically measuring hematocrit (Hct) value of blood, and has an electrode system including a working electrode and a counter electrode, and the two electrodes That is, no redox substance is disposed on the working electrode, and a redox substance is disposed on the counter electrode.
- the blood is introduced into the electrode system, and a voltage is applied to the electrode system in this state.
- the sensor detects the current value of the oxidizing current or the reducing current flowing between the electrodes when applied.
- the measuring device of the present invention is a device for measuring an Hct value, comprising: holding means for holding the sensor of the present invention; applying means for applying a constant voltage to an electrode system of the sensor; and an electrode of the sensor.
- This is a measuring device having a detecting means for detecting an oxidation current or a reduction current of the system.
- the oxidation-reduction substance is not arranged on the working electrode but is arranged on the counter electrode. Therefore, blood containing no redox substance is present on the working electrode. Therefore, in the present invention, a reliable current value depending on the Hct value of blood is obtained by the working electrode, and the current value can be detected with high sensitivity by a redox substance on the counter electrode. As a result, the measurement accuracy is excellent, and the force can be easily measured. Further, according to the present invention, the Hct value can be electrochemically measured by a sensor, so that a special large measuring instrument or device such as a conventional measuring method is used. Eliminates the need for measurement methods.
- FIG. 1 is an exploded perspective view showing an example of a sensor according to the present invention.
- FIG. 2 is a cross-sectional view of the sensor.
- FIG. 3 is a plan view of the sensor.
- FIG. 4 is an exploded perspective view showing another example of the sensor of the present invention.
- FIG. 5 is a cross-sectional view of the sensor.
- FIG. 6 is a plan view of the sensor.
- FIG. 7A is a graph showing a change with time in response current value A) to voltage application in still another example of the sensor of the present invention
- FIG. 7B is a sensitivity to voltage application in the above example. It is a graph of a change with time of a difference (%).
- FIG. 8A is a diagram showing an arrangement state of an oxidizing and reducing substance in a sensor of a comparative example
- FIG. 8B is a graph showing a change over time of a response current value (A) with respect to application of a voltage in the example
- FIG. 8C is a graph of the change over time in the sensitivity difference (%) with respect to the application of the voltage in the above example.
- FIG. 9A is a view showing an arrangement state of an oxidizing and reducing substance in a sensor of another comparative example
- FIG. 9B is a time-dependent change of a response current value (A) to application of a voltage in the example.
- FIG. 9C is a graph showing the change over time in the sensitivity difference (%) with respect to the application of the voltage in the above example.
- FIG. 10A is a view showing an arrangement state of an oxidation reducing substance in still another example of the sensor of the present invention.
- FIG. 10B is a view showing a response current value (A) to a voltage application in the example.
- FIG. 10C is a graph showing the change with time in the sensitivity difference (%) with respect to the application of the voltage in the above example.
- FIG. 11A is a view showing an arrangement state of an oxidation reducing substance in still another example of the sensor of the present invention.
- FIG. 11B is a view showing a response current value (A) to a voltage application in the example.
- FIG. 11C is a graph showing a change with time in the sensitivity difference (%) with respect to application of a voltage in the above example.
- FIG. 12A is a view showing an arrangement of an oxidizing and reducing substance in still another example of the sensor of the present invention.
- FIG. 12B is a graph showing a change in response current value (A) with time in response to voltage application in the above example
- FIG. 12C is a graph showing a difference in sensitivity to voltage application in the above example (FIG. %) Is a graph of the change over time.
- FIG. 13A is a view showing an arrangement state of an oxidation reducing substance in still another example of the sensor of the present invention.
- FIG. 13B is a view showing a response current value (A) to a voltage application in the above example.
- FIG. 13C is a graph showing the change with time in the sensitivity difference (%) with respect to the application of the voltage in the above example.
- FIG. 14A is a view showing an arrangement state of an oxidation reducing substance in still another example of the sensor of the present invention
- FIG. 14B is a view showing a response current value (A) to a voltage application in the above example.
- FIG. 14C is a graph showing the change with time in the sensitivity difference (%) with respect to the application of the voltage in the above example.
- FIG. 15A is a diagram showing an arrangement state of an oxidation reducing substance in still another example of the sensor of the present invention.
- FIG. 15B is a view showing a response current value (A) to a voltage application in the above example.
- FIG. 15C is a graph showing the change with time in the sensitivity difference (%) with respect to the application of the voltage in the above example.
- FIG. 16A is a diagram showing an arrangement state of a redox substance in a sensor of still another comparative example
- FIG. 16B is a graph showing a change with time of a response current value (A) to a voltage application in the example
- FIG. 16C is a graph showing the change with time of the sensitivity difference (%) with respect to the application of the voltage in the above example.
- FIG. 17A is a diagram showing an arrangement state of a redox substance in a sensor of still another comparative example
- FIG. 17B is a graph showing a change over time of a response current value (A) with respect to application of a voltage in the above example
- FIG. 17C is a graph showing the change over time in the sensitivity difference (%) with respect to the application of the voltage in the above example.
- FIG. 18A is a diagram showing an arrangement state of a redox substance in a sensor of still another comparative example
- FIG. 18B is a graph showing a change over time of a response current value (A) to a voltage application in the above example
- FIG. 18C is a graph of the change over time in the sensitivity difference (%) with respect to the application of the voltage in the above example.
- FIG. 19A shows the application of a voltage (0.5 V) in still another example of the sensor of the present invention.
- FIG. 19B is a graph showing the change over time in the sensitivity difference (%) to the application of the voltage in the above example.
- FIG. 20A is a graph showing a change over time in a response current value (A) of a voltage (1. OV) with respect to a marked calorie in still another example of the sensor of the present invention
- FIG. 4 is a graph showing a change with time of a sensitivity difference (%) with respect to application of a voltage.
- FIG. 21A is a graph showing a change over time in a response current value (A) with respect to application of a voltage (1.5 V) in still another example of the sensor of the present invention.
- 6 is a graph showing a change with time of a sensitivity difference (%) with respect to application of a voltage.
- FIG. 22A is a graph showing a change over time in a response current value (A) of a voltage (2. OV) with respect to a marked calorie in still another example of the sensor of the present invention
- FIG. 4 is a graph showing a change with time of a sensitivity difference (%) with respect to application of a voltage.
- FIG. 23A is a graph showing a change over time in a response current value (A) with respect to application of a voltage (2.5 V) in still another example of the sensor of the present invention.
- FIG. 6 is a graph showing a change with time of a sensitivity difference (%) with respect to application of a voltage.
- FIG. 24A is a graph showing a change over time in a response current value (A) of a voltage (3. OV) with respect to a marked calorie in still another example of the sensor of the present invention
- FIG. 4 is a graph showing a change with time of a sensitivity difference (%) with respect to application of a voltage.
- FIG. 25A is a graph showing a change over time in a response current value (A) with respect to application of a voltage (3.5 V) in still another example of the sensor of the present invention
- FIG. 6 is a graph showing a change with time of a sensitivity difference (%) with respect to application of a voltage.
- FIG. 26A is a graph showing a change over time in a response current value (A) of a voltage (4. OV) with respect to a marked amount in still another example of the sensor of the present invention.
- 4 is a graph showing a change with time of a sensitivity difference (%) with respect to application of a voltage.
- FIG. 27A is a graph showing a change over time in a response current value (A) with respect to application of a voltage (4.5 V) in still another example of the sensor of the present invention
- 6 is a graph showing a change with time of a sensitivity difference (%) with respect to application of a voltage.
- FIG. 28A is a graph showing a change over time in a response current value (A) of a voltage (5. OV) with respect to a marked calorie in still another example of the sensor of the present invention.
- A response current value
- 5. OV a voltage
- 6 is a graph of a change with time of a sensitivity difference (%) with respect to application of a voltage.
- FIG. 29A is a graph showing a change over time in a response current value (A) with respect to application of a voltage (5.5 V) in still another example of the sensor of the present invention.
- 6 is a graph showing a change with time of a sensitivity difference (%) with respect to application of a voltage.
- FIG. 30A is a graph showing a change over time of a response current value (A) of a voltage (6. OV) with respect to a marked amount in still another example of the sensor of the present invention
- FIG. 4 is a graph showing a change with time of a sensitivity difference (%) with respect to application of a voltage.
- FIG. 31A is a graph showing a change over time in a response current value (A) with respect to application of a voltage (6.5 V) in still another example of the sensor of the present invention.
- 6 is a graph showing a change with time of a sensitivity difference (%) with respect to application of a voltage.
- FIG. 32A is a graph showing a change with time in response current value (A) to voltage application in still another example of the sensor of the present invention.
- FIG. 32B is a sensitivity to voltage application in the example. It is a graph of a change with time of a difference (%).
- FIG. 33 is a perspective view showing an example of the measuring device of the present invention.
- FIG. 34 is a configuration diagram showing a configuration of the measurement apparatus of the above example.
- the redox substance is not particularly limited, and may be in a reduced state or an oxidized state.
- examples include benzoquinone, p-benzoquinone derivatives, phenazine methosulfate, methylene blue, phenol, and phenol derivatives.
- ferricyanide is preferable, and potassium ferricyanide is more preferable.
- the reduced state of the ferricyanide is ferrocyanide, and the reduced state of potassium ferricyanide is potassium ferrocyanide.
- the amount of the reducing compound is not particularly limited, and is 0.10 lOOOOmM per measurement or per sensor, and preferably 1-1500mM, more preferably. Is 10-200 mM.
- a substance which is relatively easily oxidized or reduced such as silver, copper, or silver salt, is used as an electrode material, the effects of the present invention can be obtained.
- the working electrode on which the redox substance is not disposed is coated with a polymer material for the purpose of preventing adhesion of impurities and oxidation.
- a polymer material for the purpose of preventing adhesion of impurities and oxidation.
- the polymer material include, for example, carboxymethinoresenolerose (CMC), hydroxyethynoresenorelose, hydroxypropinoresenorelose, methinoresenorelose, etinoresenorelose, etinolehydroxyethenoresorerolose, Canoleboki Polyamino acids such as cellulose cellulose, polyvinyl alcohol, polyvinylpyrrolidone, and polyzine, polystyrene sulfonic acid, gelatin and its derivatives, polyacrylic acid and its salts, polymethacrylic acid and its salts, starch and its derivatives, and maleic anhydride Heavy And its salts, agarose gels and derivatives thereof. These may
- the applied voltage between the two electrodes is preferably equal to or higher than the voltage at which water is electrolyzed, more preferably in the range of 110 V, and still more preferably. Is in the range of 1.1-6.5V.
- the hematocrit-dependent current can be measured with even higher sensitivity, and the individual is not affected by other redox substances present in the blood. A stable current independent of differences (individual differences) can be obtained.
- a negative voltage may be applied to the working electrode based on the counter electrode.
- the caloric times are, for example, 0.001 to 60 hectares, preferably 0.01 to 10 hectares, and more preferably 0.01 to 5 seconds.
- the closest distance between the working electrode and the counter electrode is 0.05 mm or more.
- the distance between the electrodes is 0.05 mm or more, the reliability of the measured value is improved. More preferably, the distance between the electrodes is at least 0.1 mm, more preferably at least 0.5 mm.
- the Hct value measurement sensor of the present invention has an insulating substrate, on which the electrode system and a flow channel for introducing the blood are formed. It is preferable that one end of the flow path communicates with the electrode system, and the other end of the flow path is opened facing the outside of the sensor, and this part serves as a blood supply port.
- the power bar may further include a spacer and a cover, and the force bar may be disposed on the insulating substrate via the spacer. /.
- a crystal homogenizing agent may be further provided in the electrode system.
- the crystal homogenizing agent is used to homogenize the crystal state of the reagent part, and includes, for example, amino acids.
- the amino acids include glycine, lanine, palin, oral isine, isoleucine, serine, threonine, methionine, asparagine, glutamine, arginine, lysine, histidine, phenylalanine, tryptophan, proline, sarcosine, and vein.
- the compounding amount of the crystal homogenizing agent is, for example, 0.1-1000 mM, preferably 10-500 mM, more preferably 10-300 mM per one measurement or one sensor.
- the voltage force applied by the application means is a voltage equal to or higher than the value at which the water is electrolyzed, and the current force Het value detected by the detection means is calculated. It is preferable to have a calculating means.
- the voltage applied is preferably in the range of 110 V, more preferably in the range of 16.5 V.
- FIG. 1, FIG. 2, and FIG. 3 show an example of the Hct value measurement sensor of the present invention.
- FIG. 1 is an exploded perspective view of the sensor
- FIG. 2 is a cross-sectional view
- FIG. 3 is a plan view
- in FIG. 1 is an exploded perspective view of the sensor
- FIG. 2 is a cross-sectional view
- FIG. 3 is a plan view
- in FIG. 1 is an exploded perspective view of the sensor
- FIG. 2 is a cross-sectional view
- FIG. 3 is a plan view
- a working electrode 11 and a counter electrode 12 are formed on an insulating substrate 101 in series.
- the surface of the working electrode 11 is preferably covered with the polymer material.
- the redox substance 13 is arranged on the counter electrode 12.
- a cover 103 is arranged on the insulating substrate 101 via a spacer 102 except for one end (the right end in the figure).
- a flow path 14 for introducing blood to the working electrode 11 and the counter electrode 12 is formed. The front end of this flow path extends to the other end of the sensor (the left end in the figure), and opens to the outside to serve as a blood supply port.
- the working electrode 11 and the counter electrode 12 are respectively connected to leads, and these leads extend to the one end side (the right side in the figure), and the tips of the leads are exposed without being covered by the cover. ing. At a portion corresponding to the end of the flow path 14 of the cover 103, an air vent hole 15 for generating a capillary phenomenon is formed.
- the material of the insulating substrate is not particularly limited, and examples thereof include polyethylene terephthalate (PET), polycarbonate (PC), polyimide (PI), polyethylene (PE), and polyethylene (PE).
- PET polyethylene terephthalate
- PC polycarbonate
- PI polyimide
- PE polyethylene
- PE polyethylene
- PE polypropylene
- PS polystyrene
- PVC polychlorinated vinyl
- POM polyoxymethylene
- MC monomer cast nylon
- PBT polybutylene terephthalate
- PMMA methacryloyl resin
- ABS resin ABS resin
- the size of the insulating substrate is not particularly limited.
- the total length is 5 to 100 mm
- the width is 3 to 50 mm
- the thickness is 0.05 to 2 mm
- the total length is 10 to 50 mm
- the width is 3 to 20 mm and the thickness is 0.1 to 1 mm. More preferably, the total length is 10 to 30 mm, the width is 3 to 10 mm, and the thickness is 0.1 to 0.6 mm.
- a conductive layer is formed by sputtering or vapor deposition using a material such as gold, platinum, or radium, and the conductive layer is formed into a specific electrode pattern by a laser.
- a laser for example, a YAG laser, a CO laser, an excimer laser, or the like can be used.
- the coating of the electrode surface with the polymer material can be formed by, for example, dissolving a predetermined polymer material in water or a buffer solution and drying it.
- a predetermined polymer material for example, 0.01 to 1.0 wt% CMC aqueous solution may be dropped on the working electrode 11 on the substrate in an amount of 0.01 mg / l and dried.
- the drying method is not particularly limited, and may be natural drying or forced drying using warm air!
- the redox substance 13 can be arranged on the counter electrode 12, for example, by dissolving the redox substance in water or a buffer solution, dropping or coating the redox substance on the surface of the counter electrode, and drying the redox substance.
- a reagent solution is prepared as described above, and the reagent solution is dropped or applied to the surface of the counter electrode and dried.
- a reagent solution is prepared by dissolving 10-200 mM potassium ferricyanide and 10-30 OmM taurine in an Owt% CMC aqueous solution, and placing the reagent solution on the counter electrode 12 on the substrate.
- One lOOmg is dropped and dried.
- the drying method is not particularly limited, and may be natural drying or forced drying using warm air.
- the material of the spacer is not particularly limited, and for example, the same material as that of the insulating substrate can be used.
- the size of the spacer is not particularly limited. In the case of the shape shown in the figure, for example, the overall length is 5 to 100 mm, the width is 3 to 50 mm, and the thickness is 0.01 to lmm. Preferably, the total length is 10 to 50 mm, the width is 3 to 20 mm, the thickness is 0.05 to 0.5 mm, and more preferably, the total length is 10 to 30 mm, the width is 3 to 10 mm, and the thickness is 0.05 to 0.25 mm. is there.
- the force at which the notch is formed in the spacer as a flow path for blood introduction The size of the force is, for example, 0.5 to 50 mm in length to the end of the blood supply rocker and 0.5 in width. 1 to 10 mm, preferably 1 to 10 mm in length to the end of the blood supply locus, 0.5 to 5 mm in width, and more preferably, 11 to 5 mm in length to the end of the blood supply locus and 0.5 to 5 in width 2 mm.
- This notch may be formed, for example, by drilling with a laser or a drill, or may be formed using a mold that can form the notch when forming the spacer. Good.
- the material of the cover is not particularly limited, and for example, the same material as that of the insulating substrate can be used. It is more preferable that the portion of the cover corresponding to the ceiling of the sample supply path is subjected to a hydrophilic treatment.
- the hydrophilic treatment include a method of applying a surfactant and a method of introducing a hydrophilic functional group such as a hydroxyl group, a carbonyl group, or a carboxyl group into the cover surface by plasma treatment or the like.
- the size of the cover is not particularly limited.
- the overall length is 5 to 100 mm
- the width is 3 to 50 mm
- the thickness is 0.01 to 0.5 mm, and preferably, the total length is 10 to 50 mm.
- the width is 3 to 20 mm
- the thickness is 0.05 to 0.25 mm, and more preferably, the total length is 15 to 30 mm
- the width is 5 to 10 mm
- the thickness is 0.05 to 0.2 mm.
- the shape in which the air vent hole is preferably formed is, for example, a circular shape, an elliptical shape, a polygonal shape, and the size thereof is, for example, a maximum diameter of 0.01 to 10 mm, preferably, The maximum diameter is 0.025-5 mm, and more preferably the maximum diameter is 0.025-2 mm. Further, even if a plurality of air vent holes are provided, no force is exerted.
- This air vent hole may be formed, for example, by drilling with a laser or a drill, or may be formed using a mold that can form an air vent portion when forming the cover.
- the senor can be manufactured by laminating an insulating substrate, a spacer, and a cover in this order, and integrally forming them.
- the three members may be attached to each other with an adhesive, or may be heat-sealed.
- an adhesive for example, an epoxy-based adhesive, an acrylic-based adhesive, a polyurethane-based adhesive, a thermosetting adhesive (such as a hot melt adhesive), a UV-curable adhesive, or the like can be used.
- the Hct value measurement using this sensor is performed, for example, as follows. That is, First, a fingertip or the like is punctured with a special lancet to bleed. On the other hand, the sensor is set in a dedicated measuring device (meter). Then, the blood supply port of the sensor set in the measuring device is brought into contact with the bleeding blood, and blood is introduced into the sensor by capillary action. By applying a constant voltage between the working electrode 11 and the counter electrode 12, the blood component is oxidized at the working electrode 11, and the oxidized reducing substance is reduced at the counter electrode 12. . The current flowing here depends on the Hct value, and this current is detected to calculate Hct.
- a calibration curve or a calibration curve table of the current and the Hct value may be prepared in advance, and converted every time the current is detected.
- the applied voltage is, for example, equal to or higher than the voltage at which water is electrolyzed, preferably 1 to 10 V, more preferably 1 to 6.5 V, and the application time is, for example, 0.001. — 60 seconds, preferably 0.01-1-10 seconds, more preferably 0.01-1-5 seconds.
- a current depending only on the Hct value of blood is generated.
- the flow and the redox substance present on the counter electrode 12 can suppress the reaction at the counter electrode 12 from becoming a rate-determining process.
- FIGS. 4, 5 and 6 show other examples of the Hct value measuring sensor of the present invention.
- FIG. 4 is an exploded perspective view of the sensor
- FIG. 5 is a sectional view
- FIG. 6 is a plan view.
- FIG. 4 is an exploded perspective view of the sensor
- FIG. 5 is a sectional view
- FIG. 6 is a plan view.
- a working electrode 21 and a counter electrode 22 are formed in parallel on a substrate 201, and an oxidation-reduction substance 23 is arranged on the counter electrode 22.
- the flow path 24 for introducing blood to the electrode also extends toward the center of the sensor at the blood introduction point at the sensor tip, but branches off into two flow paths in the middle, It is T-shaped, and a working electrode 21 or a counter electrode 22 is located at each end of the branched flow path.
- the cutout shape of the spacer 202 is also T-shaped, and air vent holes 25 for generating a capillary phenomenon are formed in portions of the cover 203 corresponding to the ends of the two branch flow paths. ing.
- Other configurations, forming materials, forming methods, Hct value measuring methods, measuring conditions, and the like are the same as those in the above-described example.
- the force exemplifying the sensor of the present invention in two examples is not limited thereto. Further, the sensor of the present invention is incorporated in a sensor for measuring blood components. It may be in a state.
- FIGS. 33 and 34 In both figures, the same parts as those in FIGS. 1 to 6 are denoted by the same reference numerals.
- FIG. 33 is a perspective view showing an example of the measuring apparatus of the present invention with a sensor mounted.
- the measuring device 123 has a sensor mounting port 125 at one end thereof, and the sensor 121 is mounted and held here.
- Reference numeral 122 denotes a sample supply port of the sensor 121.
- a display unit 124 is provided at substantially the center of the measuring device 123, and the measurement result is displayed here.
- FIG. 34 shows an example of the configuration of the measuring apparatus of the present invention.
- the measuring device 110 mainly includes two connectors 11 la and 111 b, a current-Z voltage conversion circuit 112, an AZD conversion circuit 113, a CPU 114, a liquid crystal display (LCD) 115, and a reference voltage source 116. Elements. Note that the reference voltage source 116 can be ground.
- the sensor electrode 12 is connected to a reference voltage source 116 via a connector 11 la.
- the working electrode 11 of the sensor is connected to a CPU 114 via a connector l lb, a current-Z voltage conversion circuit 112 and an AZD conversion circuit 113.
- the liquid crystal display device 115 is connected to a CPU.
- the measurement of hematocrit is performed, for example, as follows. That is, first, when blood is introduced into the electrode system of the sensor, a certain voltage is applied between the working electrode 11 and the counter electrode 12 from the current Z-voltage conversion circuit 112 and the reference voltage source 116 for a certain time in accordance with a command from the CPU 114. You. The preferable range of the applied voltage is as described above. By this voltage application, an oxidation current or a reduction current flows between the two electrodes. This current is based on the hematocrit of the blood. Then, this current is converted to a voltage by the current Z voltage conversion circuit 112, and the voltage value is converted to a digital value by the AZD conversion circuit 113 and output to the CPU 114. The CPU 114 calculates a response value based on the digital value, converts the response value into a hematocrit value, and displays the result on the liquid crystal display unit 115.
- a sensor having the configuration shown in FIGS. 1, 2 and 3 was produced.
- the working electrode 11 was covered with CMC.
- ferricyanidai potassium (amount: 60 mM)
- tau A reagent solution prepared by dissolving phosphorus (80 mM) in a CMC aqueous solution (0.1 wt%) was dropped on the counter electrode 12 and dried. The closest distance between the two electrodes was 1. Omm or more.
- three blood samples were prepared in which the Hct value was adjusted to 25, 45 and 65. For these three blood samples, the current flowing through the two electrodes of the sensor was measured by the sensor under the conditions of an applied voltage of 2.5 V and an applied time of 3 seconds. The results are shown in the graphs of FIGS.
- FIG. 7A is a graph showing the change over time of the response current value (A) with respect to the applied voltage
- FIG. 7B is a graph showing the change over time of the sensitivity difference (%) with respect to the applied voltage).
- the graph of the sensitivity difference shows the time-dependent change of the blood response value of the Hct value of 25% or 65% based on the blood response value of the Hct value of 45%.
- the difference in sensitivity did not depend on the voltage application time, and the response current reflecting the Hct value could be clearly detected. Detection was possible even when there was no polymer material such as CMC on both electrodes.
- a sensor having the configuration shown in FIG. 8A was manufactured. As shown in the figure, in this sensor, the working electrode 31 and the counter electrode 32 are formed so as to be in contact with each other on the flow path 34, and a portion corresponding to the end of the flow path 34 of the cover (not shown) has a capillary phenomenon. An air vent hole 35 for generating air is formed.
- a reagent solution was prepared by dissolving potassium ferricyanide (10-200 mM), potassium ferricyanide (about 1 / 7th the concentration of potassium ferricyanide), and taurine (10-300 mM) in 0.01% -2 Owt% CMC aqueous solution.
- FIG. 8B is a graph showing the change over time of the response current value A) with respect to the applied voltage
- FIG. 8C is a graph showing the change over time of the sensitivity difference (%) with respect to the applied voltage).
- the difference in sensitivity was greatly affected by the voltage application time, and this resulted in obtaining a response current suitable for quantifying Hct.
- Example 2 In the sensor of Comparative Example 1, 0.01% -2. A reagent solution was prepared by dissolving 10-200 mM of potassium salt and 10-300 mM of taurine, and the solution was added dropwise so as to protrude from both the working electrode 31 and the counter electrode 32 of the substrate and dried. Except for this, under the same conditions as in Example 1 (applied voltage: 2.5 V, etc.), the current flowing through the two electrodes of the sensor was measured for the sample having three Hct values. The results are shown in the graphs of FIGS. 9B and 9C. FIG. 9B is a graph showing the change over time of the response current value A) with respect to the applied voltage), and FIG. 9C is a graph showing the change over time of the sensitivity difference (%) with respect to the applied voltage). As shown in the figure, also in this comparative example, the difference in sensitivity was greatly affected by the voltage application time, and a response current suitable for quantifying Hct was obtained.
- Example 2 Example 2
- each of the sensors was produced, and in each of the sensors, the arrangement of the oxidation-reduction substance (ferricyanide potassium) with respect to the working electrode or the counter electrode was changed. Response current and sensitivity difference were measured.
- Comparative Example 3 three kinds of sensors (2-7-2-9) were produced, and in each sensor, an oxidizing and reducing substance (ferricyanidari potassium) for the working electrode or the counter electrode was prepared. The response current and the sensitivity difference were measured by changing the arrangement of.
- Each of the sensors was manufactured in the same manner as in Example 1, except for the arrangement of the redox substance and the distance between the electrodes (1.15 mm).
- FIG. A is a diagram showing the arrangement pattern of the redox substances
- FIG. B is a graph showing the change over time of the response current value (A) with respect to the applied voltage).
- Figure C is a graph of the change over time of the sensitivity difference (%) with respect to the applied voltage).
- the oxidation-reduction substance 13 is disposed so as to protrude from the counter electrode 12, and is provided on the surface of the counter electrode 12 and a part of the counter electrode between the two electrodes. And the reducing substance 13 are present.
- the results of measuring the current flowing through both electrodes of this sensor are shown in the graphs of FIGS. 10B and 10C.
- the sensitivity difference did not depend on the voltage application time, and the response current reflecting the Hct value could be clearly and satisfactorily detected.
- the redox substance 13 is disposed only on the surface of the counter electrode 12.
- the results of measuring the current flowing through both electrodes of this sensor are shown in the graphs of FIGS. 11B and 11C.
- the difference in sensitivity did not depend on the voltage application time, and the response current reflecting the Hct value could be clearly and satisfactorily detected.
- the redox substance 13 is disposed so as to protrude from the counter electrode 12, and the redox substance is present on the surface of the counter electrode 12 and between the two electrodes. Are there. Note that no redox substance exists on the working electrode 11.
- the results of measuring the current flowing through both electrodes of this sensor are shown in the graphs of FIGS. 12B and 12C. As shown in both figures, according to this sensor, the difference in sensitivity did not depend on the voltage application time, and the response current reflecting the Hct value could be clearly detected.
- the arrangement of the working electrode 11 and the counter electrode 12 is exchanged, and the counter electrode 12 in which the redox substance 13 is arranged is formed on the upstream side of the flow of the introduced blood.
- the working electrode 11 in which the oxidizing and reducing substance 13 is not disposed on the downstream side is formed.
- the results of measuring the current flowing through the two electrodes of this sensor are shown in the graphs of FIGS. 13B and 13C.
- the difference in sensitivity did not depend on the voltage application time, and the response current reflecting the Hct value could be clearly detected, but as described in (2-1) above.
- the sensitivity difference was slightly smaller than those of the examples of (2-2) and (2-3).
- the redox substance 13 is disposed so as to protrude from the counter electrode 12, and is provided on a part of the surface of the counter electrode 12 and a part between the two electrodes. There are redox substances.
- the results of measuring the current flowing through both electrodes of this sensor are shown in the graphs of FIGS. 14B and 14C.
- the sensitivity difference does not depend on the voltage application time for 1 second (between 3 and 4 seconds in the figure) immediately after the voltage is applied, and the Hct value The response current reflecting the above was clearly detected. [0047] (2-6)
- the redox substance 13 is arranged so as to protrude from the counter electrode 12, and the redox substance is present on a part of the surface of the counter electrode 12. Note that no redox substance exists between the two electrodes.
- the results of measuring the current flowing through both electrodes of this sensor are shown in the graphs of FIGS. 15B and 15C.
- the sensitivity difference does not depend on the voltage application time within 1 second (between 3 and 4 seconds in the figure) immediately after the voltage is applied. The response current reflecting the Hct value was clearly detected.
- the oxidation-reduction substance 13 is disposed on the working electrode 11, the counter electrode 12, and all of the two electrodes.
- the results of measuring the current flowing through both electrodes of this sensor are shown in the graphs of FIGS. 16B and 16C. As shown in both figures, this sensor exerted a force that could not clearly detect the response current reflecting the Hct value.
- the oxidation-reduction substance 13 is disposed on the working electrode 11, the counter electrode 12, and a part of the electrodes.
- the results of measuring the current flowing through both electrodes of this sensor are shown in the graphs of FIGS. 17B and 17C. As shown in both figures, this sensor exerted a force that could not clearly detect the response current reflecting the Hct value.
- the response current and the sensitivity difference in the sensor were measured while changing the applied voltage in the range of 0.5 to 6.5 V.
- the sensor was manufactured in the same manner as in Example 1.
- the measurement of the response current value and the sensitivity difference was also performed in the same manner as in Example 1.
- the measurement results are shown in the graphs in FIGS. Note that in Fig. 19 and Fig. 31, Fig. FIG. 6 is a graph showing a change over time in response current value (A) with respect to pressure (V), and FIG. B is a graph showing a change over time in sensitivity difference (%) with respect to applied voltage (V).
- a response current reflecting the Hct value could be detected even with an applied voltage of 0.5V, but when applied at 1-6.5V, the response current shown in Figs.
- the response current can be detected more clearly, and the most preferable case is when the voltage is applied at 11 to 13 V as shown in FIGS.
- the current based on the Hct value is detected by changing the applied voltage under a certain condition, but the present invention is not limited to this, and the applied voltage is not in the range shown in the present embodiment.
- the response current reflecting the Hct value can be clearly detected by appropriately setting other conditions such as the distance between the electrodes and the type and amount of the redox substance.
- a sensor having the configuration shown in FIGS. 1, 2 and 3 was produced.
- the working electrode 11 was covered with CMC.
- a reagent solution prepared by dissolving potassium ferrocyanide (amount: 60 mM) and taurine (80 mM) in a CMC aqueous solution (0.1 wt%) was dropped on the counter electrode 12 and dried. The closest distance between the two electrodes was 1. Omm or more.
- three blood samples were prepared in which the Hct value was adjusted to 25, 45 and 65. With respect to these three blood samples, 2.5 V was applied to the working electrode as an applied voltage by the sensor, and the current flowing through both electrodes of the sensor was measured under the condition of an application time of 3 seconds.
- FIG. 32A is a graph showing the change over time of the response current value (A) with respect to the applied voltage (V)
- FIG. 32B is a graph showing the change over time of the sensitivity difference (%) with respect to the applied voltage.
- the sensitivity difference did not depend on the voltage application time, and the response current reflecting the Hct value could be clearly detected. Detection was possible even when there was no polymer material such as CMC on both electrodes.
- the method for measuring the Hct value of the present invention and the sensor and the measuring apparatus used for the method, are capable of electrochemically measuring the Hct value with high accuracy and high reliability and easily. You can. Therefore, the measurement method, sensor, and measurement device of the present invention are useful for measuring the Hct value of blood, and are suitable for correction using the Hct value in electrochemical measurement using a sensor for blood components such as glucose.
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Abstract
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Priority Applications (7)
Application Number | Priority Date | Filing Date | Title |
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US10/578,988 US8088271B2 (en) | 2003-12-04 | 2004-12-03 | Method of measuring hematocrit (Hct), sensor used in the method, and measuring device |
EP17190588.8A EP3285068B1 (en) | 2003-12-04 | 2004-12-03 | Method of electrochemically measuring hematocrit (hct) |
EP04819923.6A EP1707953B1 (en) | 2003-12-04 | 2004-12-03 | Method of measuring hematocrit (Hct) |
CA2545477A CA2545477C (en) | 2003-12-04 | 2004-12-03 | Method of measuring hematocrit (hct), sensor used in the method, and measuring device |
JP2005515982A JP4717636B2 (ja) | 2003-12-04 | 2004-12-03 | ヘマトクリット(Hct)の測定方法およびそれに用いるセンサならびに測定装置 |
KR1020067010645A KR101049330B1 (ko) | 2003-12-04 | 2004-12-03 | 헤마토크릿의 측정 방법 및 그것에 이용하는 센서 및 측정장치 |
US13/279,787 US8480869B2 (en) | 2003-12-04 | 2011-10-24 | Method of measuring hematocrit (Hct), sensor used in the method, and measuring device |
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JP2003405481 | 2003-12-04 | ||
JP2003-405481 | 2003-12-04 |
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US10/578,988 A-371-Of-International US8088271B2 (en) | 2003-12-04 | 2004-12-03 | Method of measuring hematocrit (Hct), sensor used in the method, and measuring device |
US13/279,787 Continuation US8480869B2 (en) | 2003-12-04 | 2011-10-24 | Method of measuring hematocrit (Hct), sensor used in the method, and measuring device |
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US (2) | US8088271B2 (ja) |
EP (3) | EP3285068B1 (ja) |
JP (7) | JP4717636B2 (ja) |
KR (1) | KR101049330B1 (ja) |
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JP2011141300A (ja) | 2011-07-21 |
JP2011047964A (ja) | 2011-03-10 |
EP1707953A4 (en) | 2009-12-23 |
JP4795492B2 (ja) | 2011-10-19 |
CN100432663C (zh) | 2008-11-12 |
US20120037506A1 (en) | 2012-02-16 |
EP3285068A1 (en) | 2018-02-21 |
JP4795493B2 (ja) | 2011-10-19 |
KR101049330B1 (ko) | 2011-07-13 |
US8088271B2 (en) | 2012-01-03 |
EP2574910A1 (en) | 2013-04-03 |
JP2011141302A (ja) | 2011-07-21 |
JP4855545B2 (ja) | 2012-01-18 |
JPWO2005054839A1 (ja) | 2007-06-28 |
JP2011047962A (ja) | 2011-03-10 |
JP4794687B2 (ja) | 2011-10-19 |
JP2011047963A (ja) | 2011-03-10 |
EP3285068B1 (en) | 2020-02-12 |
EP1707953A1 (en) | 2006-10-04 |
EP2574910B1 (en) | 2017-10-25 |
KR20060113717A (ko) | 2006-11-02 |
JP4794686B2 (ja) | 2011-10-19 |
US8480869B2 (en) | 2013-07-09 |
CA2545477C (en) | 2016-06-21 |
CA2545477A1 (en) | 2005-06-16 |
US20070062822A1 (en) | 2007-03-22 |
JP2011141301A (ja) | 2011-07-21 |
CN1886652A (zh) | 2006-12-27 |
EP1707953B1 (en) | 2015-07-01 |
JP4717636B2 (ja) | 2011-07-06 |
JP4794688B2 (ja) | 2011-10-19 |
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