WO2014045584A1 - バイオセンサ及びバイオセンサの製造方法 - Google Patents
バイオセンサ及びバイオセンサの製造方法 Download PDFInfo
- Publication number
- WO2014045584A1 WO2014045584A1 PCT/JP2013/005542 JP2013005542W WO2014045584A1 WO 2014045584 A1 WO2014045584 A1 WO 2014045584A1 JP 2013005542 W JP2013005542 W JP 2013005542W WO 2014045584 A1 WO2014045584 A1 WO 2014045584A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- reagent
- electrode
- biosensor
- mediator
- biosensor according
- Prior art date
Links
Images
Classifications
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/001—Enzyme electrodes
- C12Q1/005—Enzyme electrodes involving specific analytes or enzymes
- C12Q1/006—Enzyme electrodes involving specific analytes or enzymes for glucose
-
- 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
- G01N27/3272—Test elements therefor, i.e. disposable laminated substrates with electrodes, reagent and channels
Definitions
- the present invention relates to a biosensor for measuring, for example, a blood glucose level and a method for manufacturing the biosensor.
- the configuration of this conventional biosensor has been as follows. That is, a reagent and a first electrode and a second electrode that are brought into conduction through the reagent when a biological sample is supplied to the reagent.
- the reagent includes a mediator, an oxidoreductase, and an adhesive. It was a configuration with.
- the biological sample when the biological sample is supplied to the reagent, the biological sample and the oxidoreductase react, and the mediator transmits the electrons generated at that time. At this time, the redox current flowing between the first and second electrodes is measured. Therefore, the blood glucose level was measured.
- the biosensor since the light effect reducing agent is provided on the wall surface of the storage container for storing the biosensor, the biosensor can be prevented from being deteriorated when the biosensor is stored in the storage container.
- the biosensor taken out from the storage container is immediately affected by light.
- the degree of influence can be kept to a slight level.
- deterioration becomes a problem. .
- an object of the present invention is to suppress degradation of a biosensor due to light.
- the present invention includes a first electrode, a second electrode, and a reagent part.
- the reagent part is provided in the vicinity of the first electrode and the second electrode.
- the reagent part includes a mediator, an oxidoreductase, and a substance that absorbs a light wavelength of 400 nm to 500 nm.
- the present invention includes a first electrode, a second electrode, and a reagent part.
- the reagent part includes a mediator, an oxidoreductase, and a light effect reducing agent that reduces the influence of light on the mediator.
- the present invention it is possible to provide a biosensor and a biosensor manufacturing method capable of suppressing deterioration due to light.
- the figure which shows the control block of the biological sample measuring system of FIG. (A) Exploded perspective view of the biosensor of FIG. 1, (b) Front view of the biosensor of FIG. 1 (c) Plan view of the biosensor of FIG. (A) The figure which shows the operation state in the measuring device of the biological sample measuring system of FIG. 1 (b) The figure which shows the operating state in the measuring device of the biological sample measuring system of FIG. Sectional drawing which shows the reagent part of the biosensor of FIG. The figure which shows the chemical structural formula of the blue No.
- the biological sample measurement system 100 of the present embodiment includes a biosensor 2 and a measurement device 30.
- an embodiment of the present invention is shown, and an insertion port 3 for a biosensor 2 is provided at one end of a main body case 1 constituting a measuring device 30.
- Biosensor 2 As shown in FIG. 3, the biosensor 2 is provided with three electrodes on a rectangular insulating substrate 4. The three electrodes are a blood component measurement working electrode (an example of a first electrode) 5, a blood component measurement counter electrode (an example of a second electrode) 6, and a blood component detection electrode (an example of a third electrode) 7. Are arranged opposite to each other at a predetermined interval.
- the blood component measurement working electrode 5, the blood component measurement counter electrode 6 and the blood component detection electrode 7 on one end side of the insulating substrate 4 are connected to the main body case 1 from the insertion port 3 shown in FIG. It is inserted in and is electrically connected to the measuring device by contacting the input terminal portion 8 (see FIG. 3).
- the blood component measurement working electrode 5 On the other end side of the biosensor 2 (on the side opposite to the insertion portion into the insertion port 3), the blood component measurement working electrode 5, the blood component measurement counter electrode 6 and the blood component detection electrode 7 are provided.
- the reagent part 9 is arranged.
- the reagent unit 9 includes an oxidoreductase such as glucose dehydrogenase, a mediator, and a light effect reducing agent, and optionally includes a buffer, a polymer material, an enzyme stabilizer, a crystal homogenizer, and the like as optional components. It has become.
- a cover 11 is disposed on the insulating substrate 4 and the reagent part 9 via a spacer 10.
- the blood component measurement working electrode 5, the blood component measurement counter electrode 6, and the blood component detection electrode 7 are not covered with the spacer 10 and the cover 11 and are exposed on one end side of the insulating substrate 4 (right side in FIG. 3). It is in a state.
- the biosensor 2 and the measuring device are configured such that the exposed blood component measuring working electrode 5, blood component measuring counter electrode 6 and blood component detecting electrode 7 are electrically connected to the input terminal portion 8 as described above. Has been.
- a biological sample introduction channel 12 for introducing blood is formed in the spacer 10 of the biosensor 2.
- the biological sample introduction path 12 extends from the other end side (left side in FIG. 3) of the biosensor 2 to above the reagent unit 9, and the other end side opening to the outside has a blood supply port (biological sample supply port). 13). That is, the biological sample spotted on the blood supply port 13 reaches the reagent unit 9 through the biological sample introduction path 12 by capillary action.
- the blood component measurement working electrode 5, the blood component measurement counter electrode 6 and the blood component detection electrode 7 are electrically connected to the input terminal portion 8, but specifically, the blood component
- the measurement working electrode 5 is a first input terminal (not shown) of the input terminal unit 8
- the blood component measurement counter electrode 6 is a second input terminal (not shown) of the input terminal unit 8, blood component detection.
- the pole 7 is connected to a third input terminal (not shown) of the input terminal portion 8.
- the blood component measurement counter electrode 6 is arranged closest to the blood supply port 13, followed by the blood component measurement working electrode 5, and finally the blood component detection.
- a pole 7 is arranged.
- a blood component measurement counter electrode (an example of a second electrode) 6, a blood component measurement working electrode (an example of a first electrode) 5, and a blood component detection electrode (an example of a third electrode) ) 7 is arranged.
- the cover 11 of the biosensor 2 promotes capillary action when blood is spotted on the blood supply port 13, and is a portion (blood component) beyond the blood component measuring working electrode 5 of the blood component measuring counter electrode 6.
- An air hole 14 is formed for intrusion up to the detection electrode 7).
- the material of the insulating substrate 4 is not particularly limited.
- the material of the insulating substrate 4 is not particularly limited.
- PET polyethylene terephthalate
- PC polycarbonate
- PI polyimide
- PE polyethylene
- PE polypropylene
- PS polystyrene
- PVC polyvinyl chloride
- POM polyoxymethylene
- MC monomer cast nylon
- PBT polybutylene terephthalate
- PMMA methacrylic resin
- ABS resin ABS resin
- the size of the insulating substrate 4 is not particularly limited, and is, for example, a total length of 5 to 100 mm, a width of 2 to 50 mm, and a thickness of 0.05 to 2 mm, preferably a total length of 7 to 50 mm, a width of 3 to 20 mm, and a thickness. It is 0.1 to 1 mm, and 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.
- Each electrode on the insulating substrate 4 can be formed, for example, by forming a conductive layer by sputtering or vapor deposition using gold, platinum, palladium or the like as a material, and processing this into a specific electrode pattern by a laser.
- a laser for example, a YAG laser, a CO 2 laser, an excimer laser or the like can be used.
- the electrode pattern is not limited to the one disclosed in the present invention, and any configuration may be used as long as the effect of the present invention can be realized.
- the surface of the electrode can be coated, for example, by preparing a polymer material solution, dropping or applying the solution onto the electrode surface, and then drying. Examples of drying include natural drying, air drying, hot air drying, and heat drying.
- the reagent unit 9 includes an oxidoreductase such as glucose dehydrogenase, a mediator, and a light effect reducing agent, and optionally includes a buffer, a polymer material, an enzyme stabilizer, a crystal homogenizer, and the like. It is configured to include selectively.
- an oxidoreductase such as glucose dehydrogenase, a mediator, and a light effect reducing agent
- each content has the largest water.
- water contains the most mediator in the number of moles, and the light effect reducing agent is less than the mediator.
- each content contains the most mediator in the number of moles, and the light effect reducing agent is less than the mediator.
- the mediator of the biosensor 2 to be used is not particularly limited, and examples thereof include ferricyanide, p-benzoquinone, p-benzoquinone derivatives, phenazine methosulfate, methylene blue, ferrocene, ferrocene derivatives, phenothiazine and derivatives thereof.
- the quinone compound is a compound containing quinone.
- the quinone compounds include quinones and quinone derivatives.
- Examples of the quinone derivative include compounds in which various functional groups (which may be referred to as substituents) are added to quinone.
- Examples of the quinone in the quinone compound include benzoquinone, naphthoquinone, anthraquinone, phenanthrenequinone, and phenanthroline quinone.
- Specific examples of the phenanthrenequinone include 9,10-phenanthrenequinone. More specifically, 9,10-phenanthrenequinone-2-sulfonic acid sodium (hereinafter, PQSA) may be mentioned.
- a mediator having an absorption band in a wavelength region of 400 nm to 500 nm can be mentioned.
- examples thereof include PQSA shown in FIG. 11, which will be described later, potassium ferricyanide, ferrocenylmethyldodecyldimethylammonium bromide, hexachlorooxamate, and the like shown in FIGS. 17 (a) to 17 (c).
- the blending amount of the mediator is not particularly limited, and is, for example, 0.1 to 1000 mM, preferably 1 to 500 mM, more preferably 10 to 200 mM per measurement or per biosensor. .
- Oxidoreductase examples include glucose oxidase, lactate oxidase, cholesterol oxidase, bilirubin oxidase, glucose dehydrogenase, and lactate dehydrogenase.
- the amount of oxidoreductase is, for example, 0.01 to 100 U, preferably 0.05 to 10 U, more preferably 0.1 to 100 U per sensor or per measurement. 5U.
- glucose it is preferable to use glucose as a measurement target, and in this case, oxidoreductase is preferably glucose oxidase and glucose dehydrogenase.
- the reagent unit 9 includes an oxidoreductase such as glucose dehydrogenase, a mediator, and a light effect reducing agent, and a buffer, a polymer material, an enzyme stabilizer, a crystal homogenizer, and the like are selected as optional components.
- an oxidoreductase such as glucose dehydrogenase, a mediator, and a light effect reducing agent
- a buffer, a polymer material, an enzyme stabilizer, a crystal homogenizer, and the like are selected as optional components.
- each of these substances is dissolved in approximately 80% of the total water.
- the reagent solution in which each substance is dissolved is used as a blood component measurement counter electrode (an example of the second electrode) 6, a blood component measurement working electrode (an example of the first electrode) 5, and a blood component detection electrode (the third electrode) of the insulating substrate 4.
- the reagent part 9 can be formed by dropping on the example 7) and drying it.
- the moisture is the most, then the mediator, then the light effect reducing agent, and then a lot of other substances are contained, but in the dry state, all the water evaporates,
- the mediator is the most, followed by the light effect mitigating agent, followed by a large amount of other substances.
- the material of the spacer 10 is not particularly limited, and for example, the same material as that of the insulating substrate 4 can be used.
- the size of the spacer 10 (including an adhesive layer such as hot melt) is not particularly limited, and is, for example, a total length of 5 to 100 mm, a width of 3 to 50 mm, and a thickness of 0.01 to 1 mm. The thickness is 7 to 50 mm, the width is 3 to 20 mm, and the thickness is 0.05 to 0.5 mm. 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.
- the spacer 10 is formed with an I-shaped notch that becomes a biological sample introduction path 12 for blood introduction.
- the material of the cover 11 is not particularly limited, and for example, the same material as that of the insulating substrate 4 can be used. More preferably, the portion of the cover 11 corresponding to the ceiling portion of the biological sample introduction path 12 is subjected to hydrophilic treatment.
- the hydrophilic treatment include a method of applying a surfactant or a method of introducing a hydrophilic functional group such as a hydroxyl group, a carbonyl group, or a carboxyl group into the surface of the cover 11 by plasma treatment or the like.
- the size of the cover 11 is not particularly limited, and is, for example, a total length of 5 to 100 mm, a width of 3 to 50 mm, and a thickness of 0.01 to 0.5 mm, preferably a total length of 10 to 50 mm, a width of 3 to 20 mm, and a thickness of 0 .05 to 0.25 mm, more preferably 15 to 30 mm in total length, 3 to 10 mm in width, and 0.05 to 0.1 mm in thickness.
- Air holes 14 are preferably formed in the cover 11, and the shape is, for example, a circle, an ellipse, or a polygon, and the size is, for example, a maximum diameter of 0.01 to 10 mm, preferably The maximum diameter is 0.05 to 5 mm, more preferably the maximum diameter is 0.1 to 2 mm.
- the air holes 14 may be formed by, for example, drilling with a laser, a drill, or the like, or may be formed using a mold that can form an air vent when the cover 11 is formed.
- the biosensor 2 can be manufactured by laminating the insulating substrate 4, the spacer 10, and the cover 11 in this order and integrating them.
- the three members may be attached with an adhesive or may be heat-sealed.
- the adhesive examples include epoxy adhesives, acrylic adhesives, polyurethane adhesives, thermosetting adhesives (hot melt adhesives, etc.), UV curable adhesives, and the like.
- the cover 11 is formed to be transparent or translucent so that the blood entering the biological sample introduction path 12 can be visually observed.
- the measuring device 30 has the main body case 1, and the main body case 1 is formed with the insertion port 3 for the biosensor 2 (see FIG. 1). An input terminal portion 8 that comes into contact with the biosensor 2 is provided at the back of the insertion port 3.
- a voltage application unit 15 for applying a voltage and a current-voltage conversion unit 16 are connected to the input terminal unit 8 via a switching circuit 17.
- the application voltage unit 18 of the voltage application unit 15 includes a switching circuit 17, an input terminal unit 8, a blood component measurement working electrode 5 of the biosensor 2, a reagent unit 9, a blood component measurement counter electrode 6, and a blood component detection electrode. 7 and the reference voltage unit 19 of the voltage application unit 15.
- the voltage of the applied voltage unit 18 is 300 mV and the voltage of the reference voltage unit 19 is 200 mV, a voltage of 100 mV is applied between the blood component measurement working electrode 5 and the blood component measurement counter electrode 6. It becomes a state.
- the voltage of the reference voltage unit 19 is constant, and the voltage waveform is obtained by changing the voltage of the applied voltage unit 18.
- the reason for providing the reference voltage unit 19 is to reduce the influence of noise on the supply voltage from the power supply unit 20.
- a current based on the potential difference between the blood component measuring working electrode 5 and the blood component measuring counter electrode 6 flows between the blood component measuring working electrode 5 and the blood component measuring counter electrode 6, and the current flowing there is converted into a voltage.
- this is expressed as a current-voltage converter (for example, a resistor provided between blood component measuring counter electrode 6 and reference voltage unit 19).
- a voltage is applied to the voltage application unit 15 via the control unit 21, and the current flowing through the biosensor 2 at that time is converted into a voltage by the current-voltage conversion unit 16. Thereafter, this voltage is digitally converted by the A / D converter 22, and the digitally converted voltage is compared with a threshold value by the determination means 23.
- the glucose level detected by the biosensor 2 and the determination result by the determination unit 23 are displayed on the display unit 24 connected to the control unit 21.
- the power supply unit 20 in FIG. 2 is for supplying power to the respective units.
- Reference numeral 25 denotes a memory unit including a table including a hematocrit value, an applied voltage at the time of glucose measurement, an application time, and the like, and a calibration curve and a calibration table created in advance from the environmental temperature.
- a clock 26 is connected to the control unit 21, and the control unit 21 is configured to execute various control operations using the time and time of the clock 26. Furthermore, a correction means 27 is provided in the control unit 21 to correct the measured blood glucose level in consideration of the influence of hematocrit value and various interfering substances, thereby increasing the measurement accuracy of the blood glucose level.
- the input terminal portion 8 is connected to the blood component measurement working electrode 5, blood component measurement counter electrode 6, and blood component detection electrode 7.
- a switch (not shown) in the insertion port 3 is pressed by the biosensor 2.
- the control unit 21 determines that the biosensor 2 is attached, and puts the measuring device 30 in the sample standby state.
- the applied voltage unit 18 starts voltage application between the blood component measurement working electrode 5 and the blood component detection electrode 7 via the input terminal unit 8, and the current -After the voltage converter 16 starts current measurement, the liquid sample has not yet been measured.
- the liquid sample is drawn from the blood supply port 13 into the biological sample introduction path 12 by capillary action.
- a liquid sample derived from a living body such as blood, sweat or urine
- a liquid sample derived from the environment a liquid sample derived from food, or the like
- the biosensor 2 is used as a blood glucose level sensor
- the user punctures his / her finger, palm, arm, etc., squeezes a small amount of blood, and uses the blood as a liquid sample to measure with the biosensor 2.
- the biosensor 2 is used as a blood glucose level sensor
- the control unit 21 determines that the liquid sample has been sucked into the biosensor 2. When the suction of the liquid sample is detected in this way, the measurement is started.
- components such as enzymes and mediators in the reagent unit 9 are dissolved in the liquid sample.
- the liquid sample, the enzyme, and the mediator come into contact with each other on the blood component measurement working electrode 5 and the blood component measurement counter electrode 6 of the biosensor 2.
- the switching circuit 17 is connected to the reference voltage unit 19 and the current-voltage conversion unit 16 under the control of the control unit 21.
- a voltage is applied between the blood component measurement working electrode 5 and the blood component measurement counter electrode 6.
- the voltage applied here will be described in detail below with reference to FIGS. 4A and 4B, but the blood component measurement working electrode 5 and the blood component measurement counter electrode in the second application period described later. 6 is transmitted to the current-voltage converter 16.
- the current that has flowed to the current-voltage converter 16 is converted into a voltage.
- This voltage is further converted into a pulse by the A / D converter 22.
- the control unit 21 performs correction by the correction unit 27 and calculates the concentration of the specific component.
- the value calculated by the control unit 21 is displayed on the display unit 24. At this time, other information to the user may be displayed together.
- FIG. 4A shows the applied voltage.
- FIG. 4B is a diagram illustrating a signal generated by applying a voltage.
- FIG. 4A two application patterns of a first application period and a second application period are shown with a pause time (for example, 1 second) interposed therebetween.
- the first application period is also referred to as a pretreatment voltage application mode or a pre-application mode.
- an applied voltage shown in FIG. 4A is provided between the blood component measurement working electrode 5 and the blood component measurement counter electrode 6 via the input terminal unit 8 in accordance with an instruction from the control unit 21.
- V1 for example, 0.35 V
- T1 for example, 2 seconds
- the rest period (rest mode) is applied between the blood component measurement working electrode 5 and the blood component measurement counter electrode 6 via the input terminal portion 8 after the pretreatment voltage application period (first application period).
- This is a period during which the pretreatment voltage application is stopped. That is, glucose in blood and oxidoreductase are allowed to react for a certain period of time.
- the second application period is a measurement voltage application mode, and after the pause period, between the blood component measurement working electrode 5 and the blood component measurement counter electrode 6 via the input terminal portion 8, FIG. Is a period during which biological information such as blood glucose level is measured by applying an application voltage T2 (for example, 0.25 V) for an application time T2 (for example, 2 seconds 3 seconds after the start of measurement).
- an application voltage T2 for example, 0.25 V
- an application time T2 for example, 2 seconds 3 seconds after the start of measurement.
- the blood component measurement working electrode 5 and the blood component measurement counter electrode 6 are obtained by enzymatic reaction.
- the reduced state electron carrier (mediator) generated on the substrate is oxidized, and its oxidation current is detected.
- the oxidation current detected by the biological sample measurement system 100 is output as a signal waveform as shown in FIG.
- target biological information such as a blood glucose level that is a glucose concentration
- T1 and T2 the application times
- the applied voltage V1 is higher than the applied voltage V2.
- the present invention is not limited to this, and the applied voltage V1 is more than the applied voltage V2.
- the applied voltage V1 and the applied voltage V2 may be the same.
- the most characteristic feature of the present embodiment is that a light effect reducing agent is mixed in the reagent unit 9.
- the reagent portion 9 has a circular shape in plan view, and when it is cut at the central portion, it is in the state shown in FIG. That is, the reagent part 9 is formed in a substantially cylindrical shape.
- the reagent part 9 is in a state in which the thickness d2 of the outer peripheral part is thicker than the thickness d1 of the inner peripheral part in a plan view on the insulating substrate 4.
- the thickness d2 of the outer peripheral part is made thicker than the thickness d1 of the inner peripheral part, which can be easily realized at the time of manufacture. I can do it.
- the reagent part 9 contains an oxidoreductase such as glucose dehydrogenase, a mediator, and a light effect reducing agent in approximately 80% of the total water, and as an optional component, a buffer, a polymer A reagent solution in a state of selectively containing materials, enzyme stabilizers, crystal homogenizing agents, and the like is dropped on the insulating substrate 4 and then dried.
- an oxidoreductase such as glucose dehydrogenase, a mediator, and a light effect reducing agent in approximately 80% of the total water
- the reagent part 9 in the dry state has a thickness d2 of the outer peripheral part thicker than the thickness d1 of the inner peripheral part in a plan view on the insulating substrate 4.
- the light effect reducing agent is more unevenly distributed on the surface portion (on the cover 11 side) than inside the reagent portion 9.
- Blue No. 1 (Brilliant Blue FCF), Green No. 3 (Fast Green FCF), Yellow No. 4 (Tartrazine) and the like can be used.
- FIG. 6 shows a chemical structural formula of Blue No. 1 (Brilliant Blue FCF) as a light effect reducing agent
- FIG. 7 shows a chemical structural formula of Green No. 3 (Fast Green FCF)
- FIG. The chemical structural formula of No. (Tartrazine) is shown.
- These Blue No. 1, Green No. 3 and Yellow No. 4 all have a hydrophobic benzene ring portion and a hydrophilic benzene ring portion as chemical structural formulas.
- the benzene ring portion includes a benzene ring substituent, a reduced form, a derivative, and the like, and may have two or more substituents.
- the hydrophilic benzene ring portion indicates a sulfophenyl group portion in the chemical structural formula
- the hydrophobic benzene ring portion indicates 4-ethyl in the chemical structural formula.
- a part of aminophenyl-methylidenylcyclohexadiene ( ⁇ part surrounded by a dotted line) is shown.
- the hydrophilic benzene ring portion indicates a sulfophenyl group or a 4-hydroxy-2-sulfophenyl group in the chemical structural formula.
- FIG. 9 is a schematic diagram showing a reagent in a state where Blue No. 1 is not added and a reagent in a state where Blue No. 1 is added. As shown in FIG. 9, in the reagent part 9 to which the blue No. 1 is added, the blue No. 1 as an example of the light effect reducing agent is unevenly distributed on the surface of the reagent part 9.
- This state is also expressed as a state in which the hydrophobic benzene ring part of the light effect reducing agent is arranged toward the surface layer side of the reagent part 9. This is extremely important, and as described above, the light effect reducing agent is more unevenly distributed in the surface portion of the reagent part 9 than in the inside, even though only a small amount is mixed with the mediator. As a result, the influence of light on the reagent part 9 can be effectively reduced.
- the reagent unit 9 is affected by light, and particularly the function of the mediator is deteriorated. Therefore, the suppression of this is the biggest feature point in the present embodiment.
- the mediator of the reagent unit 9 used in this embodiment is affected by light of, for example, 400 to 500 nm
- the light effect reducing agent to be used is blue No. 1 (brilliant blue FCF), green as described above. No. 3 (Fast Green FCF) or Yellow No. 4 (Tartrazine).
- Blue No. 1 (Brilliant Blue FCF), Green No. 3 (Fast Green FCF) and Yellow No. 4 (Tartrazine) absorb 400-500 nm light and degrade themselves (for example, it becomes lighter in color) However, it is difficult for the light of 400 to 500 nm to reach the mediator, and as a result, the function of the mediator is not easily deteriorated.
- the electron transfer function by the mediator is appropriately exhibited, and as a result, the measurement of an appropriate biological sample is possible. Will be done.
- FIG. 11 (a) is a graph showing absorption spectra of Green No. 3 and PQSA around 350 nm to 500 nm.
- FIG. 11B is a diagram showing absorption spectra of Blue No. 1 and PQSA in the vicinity of 350 nm to 500 nm.
- FIG. 11 (c) is a graph showing absorption spectra of Yellow No. 4 and PQSA around 350 nm to 500 nm.
- the concentration of Green No. 3 is 0.5 mM
- the concentration of PQSA is 0.1 mM.
- concentration of blue No. 1 is 0.1 mM, and the density
- concentration of yellow No. 4 is 0.01 mM and the density
- green No. 3, blue No. 1 and yellow No. 4 absorb absorption in the vicinity of the wavelength (414 nm) affecting PQSA.
- Green No. 3, Blue No. 1 and Yellow No. 4 have a ⁇ ratio higher than at least 1 near the wavelength (414 nm), and thus absorb light more easily than PQSA. Deterioration due to light absorption can be prevented.
- FIG. 12 is a process diagram showing a method for manufacturing a biosensor.
- the insulating substrate 4 on which the blood component measurement working electrode 5, the blood component measurement counter electrode 6 and the blood component detection electrode 7 are formed is produced.
- the reagent solution is adjusted.
- As the reagent solution at least an oxidoreductase that oxidizes or dehydrogenates a measurement target, a mediator that exchanges electrons with the oxidoreductase, and a light effect reducing agent are dissolved in water. It is prepared by selectively dissolving molecular materials, enzyme stabilizers, crystal homogenizers, and the like.
- the reagent solution is applied to the blood component measuring working electrode 5, the blood component measuring counter electrode 6 and the blood component detecting electrode 7 on the insulating substrate 4 on which each electrode is formed. It is dripped (an example of a mounting process).
- the reagent solution placed on the insulating substrate 4 is dried (an example of a drying step).
- the insulating substrate 4, the spacer 10, and the cover 11 are stacked in this order and integrated to manufacture a biosensor.
- FIG. 13A is a diagram showing a graph of the sunlight exposure effect when Green 3 is used.
- FIG.13 (b) is a figure which shows the graph of the sunlight exposure effect at the time of using blue No. 1.
- FIG. 13 (c) is a figure which shows the graph of the sunlight exposure influence at the time of using yellow No. 4.
- FIG. 13A is a diagram showing a graph of the sunlight exposure effect when Green 3 is used.
- FIG.13 (b) is a figure which shows the graph of the sunlight exposure effect at the time of using blue No. 1.
- FIG. 13 (c) is a figure which shows the graph of the sunlight exposure influence at the time of using yellow No. 4.
- a reagent solution is prepared by adding the reagent shown in FIG. 14A to water to prepare a reagent solution, and then drying the reagent solution.
- the composition of the reagent solution is 91.25% in wt (wt)%, 7.41% glucose dehydrogenase as an enzyme, 0.08% green 3 (molecular weight 808.87), K 2 HPO 4 , KH 2 PO 4 and NaH as buffers.
- a polymer material is CMC (carboxymethylcellulose) is 0.13% as an adhesive
- ATA enzyme stabilizers (3-amino-1,2,4-triazole)
- Maltitol is 0.17% as a stabilizer
- PQSA is 0.70% as a mediator
- other selection members are 0.03%.
- a reagent solution having a concentration of 10 mM in green 3 is prepared with the composition shown in FIG.
- the concentration of PQSA molecular weight 309.28
- the concentration of PQSA is 22.6 mM in any case.
- a reagent solution to which no dye is added is also prepared as a comparative example.
- a reagent part was created using these three types of reagent solutions, and the influence of the measured values due to exposure to sunlight was confirmed.
- Green No. 3 1 mM reagent solution, and Green No. 3 10 mM reagent container a reagent part is prepared, and the reagent part is used for exposure to sunlight for 1 hour. If not, compare the measurement results of the sample solution.
- FIGS. 14A and 14B show the weight (wt)% of each component of the reagent part after drying.
- the average value of five measurements when measuring a sample solution is 41.5 mg /
- the sample solution of Hct42% having a glucose concentration of about 48 mg / dL was measured using a dye-free reagent after exposure to sunlight, the average value of 5 measurements was 87.2 mg / dL.
- the divergence rate in the graph of FIG. 13 is obtained by the equation of (measured value after exposure to sunlight ⁇ measured value without exposure to sunlight) / measured value without exposure to sunlight.
- the average value of 5 times was 41.7 mg / dL
- the above sample was obtained using the green No. 3 1 mM reagent with sunlight exposure.
- the average value of 5 times was 76.4 mg / dL.
- FIG. 13B (Effect of exposure to sunlight from Blue No. 1)
- the reagent shown in FIG. 15A is added to water to prepare a reagent solution, and the reagent solution is dried to prepare a measurement reagent.
- Blue No. 1 in the reagent solution for forming the reagent part 9 is 1 mM or more, the effect of suppressing the exposure to sunlight is exerted.
- FIG. 13 (b) it can be seen that when Blue No. 1 uses a reagent solution having a concentration of 5 mM, an inhibitory effect is obtained. Therefore, Blue No. 1 in the reagent solution for forming the reagent portion 9 is obtained.
- the concentration is more preferably in the range of 1 mM or more and 5 mM or less.
- FIG. 13C for example, the reagent shown in FIG. 16A was added to water to prepare a solution, and the solution was dried to prepare a measurement reagent.
- a reagent part was prepared using each of the yellow No. 4 1 mM reagent solution, the yellow No. 4 5 mM reagent solution, and the yellow No. 4 10 mM reagent solution, and the reagent part was used to expose to sunlight for 1 hour. When not exposed, the measurement results of the sample solutions similar to those of Green No. 3 are compared. 16A to 16C show the weight (wt)% of each component of the reagent part after drying.
- the divergence rate when each reagent part is used by the same calculation as that of Green No. 3 is 66.3% when Yellow No. 4 is 1 mM, and the deviation rate is 28.4 when Yellow No. 4 is 5 mM. In the case of Yellow No. 4 10 mM, the deviation rate was 19.5%.
- each biosensor 2 itself can suppress deterioration due to light.
- the conventional product there is a case where the actual use becomes impossible due to the influence of light. For example, even if the biosensor 2 taken out from the container is left for one hour, the conventional product (light Compared with no effect reducing agent), it is less affected by light and can be used as it is.
- the biosensor 2 of the above embodiment includes a blood component measurement working electrode 5, a blood component measurement counter electrode 6, and a reagent unit 9.
- the reagent part 9 is provided in the vicinity of the blood component measurement working electrode 5 and the blood component measurement counter electrode 6.
- the reagent part 9 includes a mediator, an oxidoreductase, and a substance that absorbs a light wavelength of 400 nm to 500 nm.
- the biosensor 2 of the above embodiment includes a blood component measurement working electrode 5, a blood component measurement counter electrode 6, and a reagent unit 9.
- the reagent part 9 is provided in the vicinity of the blood component measurement working electrode 5 and the blood component measurement counter electrode 6.
- the reagent part 9 includes a mediator, an oxidoreductase, and a light effect reducing agent.
- the biosensor 2 of the above embodiment includes an insulating substrate 4 (an example of a substrate) on which at least one of the blood component measurement working electrode 5 and the blood component measurement counter electrode 6 is disposed.
- the reagent part 9 is disposed on at least one electrode of the insulating substrate 4.
- the reagent portion 9 is arranged on the insulating substrate 4, and the outer peripheral thickness d2 is larger than the inner thickness d1. It is formed thick.
- the biosensor 2 of the above embodiment includes a spacer 10 and a cover 11 as shown in FIG.
- the spacer 10 is laminated on the side of the insulating substrate 4 on which the reagent part 9 is formed.
- the cover 11 is laminated on the side of the spacer 10 opposite to the insulating substrate 4 and is transparent or translucent.
- the spacer 10 is formed with a biological sample introduction part 12 that guides the biological sample to the reagent part 9 at a part corresponding to the reagent part 9.
- the concentration of the measurement object can be measured with a small amount of biological sample.
- the substance that absorbs the light wavelength of 400 nm to 500 nm has a hydrophobic benzene ring portion and a hydrophilic benzene ring portion in the chemical structural formula.
- the hydrophobic benzene ring part is exposed on the surface side (the cover 11 side), that is, in the situation where it floats to the surface side of the reagent part 9. Drying proceeds, and as a result, a substance that absorbs a light wavelength of 400 nm to 500 nm is more unevenly distributed on the surface portion (cover 11 side) than in the reagent portion 9. Thereby, the influence of the light with respect to a mediator can be effectively reduced by a small amount of light influence reducing agent.
- the substance that absorbs the light wavelength of 400 nm to 500 nm is arranged with the hydrophobic benzene ring portion facing the surface layer side of the reagent portion.
- the influence of light on the mediator can be effectively reduced by a substance that absorbs a small amount of light wavelength of 400 nm to 500 nm.
- the substance that absorbs the light wavelength of 400 nm to 500 nm is Brilliant Blue FCF (Blue No. 1), Fast Green FCF (Green No. 3), or Tartrazine (Yellow No. 4).
- the influence of light on the mediator can be effectively reduced by a substance that absorbs a light wavelength of 400 nm to 500 nm that is smaller than that of the mediator.
- the weight ratio of the fast green FCF to the entire reagent part 9 is 6.01% or more and 39.02% or less.
- the weight ratio of the fast green FCF to the entire reagent part 9 is 11.6% to 116% of the weight ratio of the mediator to the entire reagent part 9.
- the weight ratio of the brilliant blue FCF to the entire reagent part 9 is 5.9% or more and 23.87% or less.
- the weight ratio of tartrazine to the entire reagent part 9 is 7.66% or more and 76.6% or less of the weight ratio of the mediator to the entire reagent part 9.
- the light effect reducing agent is a substance that absorbs a light wavelength of 400 nm to 500 nm.
- the manufacturing method of a biosensor is provided with the mounting process and the drying process.
- a reagent solution containing at least an oxidoreductase, a mediator, and fast green FCF is placed on the first electrode and the second electrode formed on the substrate.
- the placed reagent solution is dried.
- the concentration of Fast Green FCF in the reagent solution is 1 mM or more and 10 mM or less.
- the manufacturing method of a biosensor is provided with the mounting process and the drying process.
- a reagent solution containing at least an oxidoreductase, a mediator, and a brilliant blue FCF is placed on the first electrode and the second electrode formed on the substrate.
- the placed reagent solution is dried.
- the concentration of brilliant blue FCF in the reagent solution is 1 mM or more and 5 mM or less.
- FIG. 17 (a), FIG. 17 (b), and FIG. 17 (c) are diagrams showing absorption spectra of potassium ferricyanide, ferrocenylmethyldodecylammonium bromide, and hexachlorooxamate used as mediators, respectively.
- the light effect reducing agent of the present embodiment it is possible to reduce the effect on the measurement value due to exposure to sunlight. That is, the light effect reducing agent of the present invention can be more effective when using a mediator that absorbs light having a wavelength in the vicinity of 400 to 500 nm.
- composition of the reagent of the present invention is not limited to the composition used in the above embodiment.
- K 2 HPO 4 0.0016 g, KH 2 PO 4 and NaH 2 PO 4 are used as the buffering agent, but not limited to these, other buffering agents such as HEPES are used. May be.
- each biosensor itself can be made less susceptible to light, and as a result, deterioration of the biosensor can be suppressed.
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Biochemistry (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biophysics (AREA)
- Engineering & Computer Science (AREA)
- Electrochemistry (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Hematology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- Emergency Medicine (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Description
すなわち、試薬と、この試薬に対する生体試料の供給時に、この試薬を介して導通状態になる第1、第2の電極とを備え、前記試薬は、メディエータと、酸化還元酵素と、接着剤とを備えた構成となっていた。
このため、従来は、バイオセンサを収容する収容容器の壁面に光影響軽減剤を設ける対応も図られている(たとえば、特許文献1)。
勿論、収容容器から取り出したバイオセンサを直ちに使用する場合には、その影響度合も軽微なものに留めることが出来るが、使用までの間に長時間放置された場合には、劣化が問題となる。
そこで本発明は、バイオセンサの光による劣化を抑制することを目的とするものである。
(実施の形態1)
本発明にかかる実施の形態の生体試料測定システム100について以下に説明する。
図1に示すように、本実施の形態の生体試料測定システム100は、バイオセンサ2と、測定装置30を備えている。
(バイオセンサ2)
バイオセンサ2には、図3に示すように、長方形状である絶縁基板4の上に、3個の電極が設けられている。3個の電極は、血液成分測定作用極(第1の電極の一例)5、血液成分測定対極(第2の電極の一例)6及び血液成分検知極(第3の電極の一例)7であり、所定の間隔を置いて対向配置されている。
(絶縁基板4)
絶縁基板4の材質は特に制限されず、例えば、ポリエチレンテレフタレート(PET)、ポリカーボネート(PC)、ポリイミド(PI)、ポリエチレン(PE)、ポリプロピレン(PP)、ポリスチレン(PS)、ポリ塩化ビニル(PVC)、ポリオキシメチレン(POM)、モノマーキャストナイロン(MC)、ポリブチレンテレフタレート(PBT)、メタクリル樹脂(PMMA)、ABS樹脂(ABS)、ガラス等が使用できる。このなかで、ポリエチレンテレフタレート(PET)、ポリカーボネート(PC)、ポリイミド(PI)が好ましく、より好ましくは、ポリエチレンテレフタレート(PET)である。
試薬部9は、上述のように、グルコースデヒドロゲナーゼ等の酸化還元酵素、メディエータ、光影響軽減剤を、含み、任意成分として、緩衝剤、高分子材料、酵素安定化剤、結晶均質化剤等を選択的に含む構成となっている。
使用されるバイオセンサ2のメディエータは、特に制限されず、例えば、フェリシアン化物、p-ベンゾキノン、p-ベンゾキノン誘導体、フェナジンメトサルフェート、メチレンブルー、フェロセン、フェロセン誘導体、フェノチアジンおよびその誘導体などが挙げられる。
酸化還元酵素としては、例えば、グルコースオキシダーゼ、ラクテートオキシダーゼ、コレステロールオキシダーゼ、ビリルビンオキシダーゼ、グルコースデヒドロゲナーゼ、ラクテートデヒドロゲナーゼなどがある。酸化還元酵素の量は、例えば、センサ1個当り、もしくは1回の測定当り、例えば、0.01~100Uであり、好ましくは、0.05~10Uであり、より好ましくは、0.1~5Uである。このなかでも、グルコースを測定対象にすることが好ましく、この場合の酸化還元酵素は、グルコースオキシダーゼおよびグルコースデヒドロゲナーゼが好ましい。
次に、スペーサー10の材質は、特に制限されず、例えば、絶縁基板4と同様の材料が使用できる。また、スペーサー10の大きさ(ホットメルト等の接着剤層も含む)は、特に制限されず、例えば、全長5~100mm、幅3~50mm、厚み0.01~1mmであり、好ましくは、全長7~50mm、幅3~20mm、厚み0.05~0.5mmであり、より好ましくは、全長10~30mm、幅3~10mm、厚み0.05~0.25mmである。スペーサー10には、血液導入のための生体試料導入路12となるI字形状の切欠部が形成されている。
また、カバー11の材質は、特に制限されず、例えば、絶縁基板4と同様の材料が使用できる。カバー11の生体試料導入路12の天井部に相当する部分は、親水性処理することが、更に好ましい。親水性処理としては、例えば、界面活性剤を塗布する方法、又はプラズマ処理などによりカバー11の表面に水酸基、カルボニル基、カルボキシル基などの親水性官能基を導入する方法がある。
(測定装置30)
上述したように、測定装置30は、本体ケース1を有し、本体ケース1には、バイオセンサ2の挿入口3が形成されている(図1参照)。そして、挿入口3の奥には、バイオセンサ2と接触する入力端子部8が設けられている。
具体的には、電圧印加部15の印加電圧部18は、切替回路17、入力端子部8、バイオセンサ2の血液成分測定作用極5、試薬部9、血液成分測定対極6、血液成分検知極7、及び電圧印加部15の基準電圧部19へと接続されている。
なお、基準電圧部19を設ける理由は、電源部20からの供給電圧のノイズによる影響を小さくするためである。
なお、図2の電源部20は、前記各部に電源を供給するためのものである。
さらに、制御部21内には、補正手段27が設けられ、測定した血糖値をヘマトクリット値や各種妨害物質の影響を考慮して補正することで、血糖値の測定精度を高めるものである。
バイオセンサ2が挿入口3に差し込まれると、入力端子部8が、血液成分測定作用極5、血液成分測定対極6及び血液成分検知極7に接続される。また、挿入口3内のスイッチ(図示せず)がバイオセンサ2に押下される。スイッチの押下により、制御部21はバイオセンサ2が装着されたと判断し、測定器30を試料待機状態とする。試料待機状態とは、制御部21の制御の下、印加電圧部18が入力端子部8を介して、血液成分測定作用極5及び血液成分検知極7間への電圧印加を開始し、かつ電流―電圧変換部16が電流測定を開始した後であって、液体試料がまだ測定に供されていない状態である。
液体試料としては、例えば、血液、汗、尿等の生体由来の液体試料や、環境由来の液体試料、食品由来の液体試料等が用いられる。例えば、バイオセンサ2を血糖値センサとして用いる場合、使用者は、自身の指、掌、又は腕等を穿刺して、少量の血液を搾り出し、この血液を液体試料として、バイオセンサ2での測定に供する。
(測定における印加電圧)
次に、測定における印加電圧について、図4(a)及び図4(b)を用いて説明する。図4(a)は、印加電圧を示す図である。図4(b)は、電圧を印加することによって生じる信号を示す図である。
第1の印加期間は、前処理電圧印加モードまたはプリ印加モードとも言われている。この第1の印加期間は、制御部21からの指示により、入力端子部8を経由して、血液成分測定作用極5と血液成分測定対極6の間に、図4(a)に示す印加電圧V1(例えば、0.35V)を印加時間T1(例えば、2秒間)、前処理電圧として印加する期間である。
ここで、図4(a)では、印加時間T1とT2の間に印加しない休止期間を設けているが、本発明は、これに限られない。
以上の構成において、本実施の形態における最も特徴的な事は、試薬部9に光影響軽減剤を混入させたことである。
このように、試薬部9を、絶縁基板4上における平面視の状態で、内周部分の肉厚d1よりも外周部分の肉厚d2を厚くする事は、その製造時において簡単に実現することが出来る。
そして、このような乾燥状態においては、光影響軽減剤は、試薬部9の内部よりも表面部分(カバー11側)に多く偏在した状態となる。
この事は、極めて重要なこととなり、上述のように、光影響軽減剤は、メディエータよりも少量しか混入させていないにもかかわらず、試薬部9の表面部分においては、内部よりも多く偏在することで、この試薬部9に対する光の影響を効果的に軽減することが出来るものとなる。
図10は、青色1号、緑色3号及び黄色4号のそれぞれの最大吸収波長(nm)、その波長における吸光度(Abs)、その吸光度を示す濃度(c(M))、モル吸光係数(ε)、及びメディエータの一例であるPQSAとのモル吸光係数との比を、PQSAとともに示す表である。尚、Abs=εcl(lはセル長であり、1cmに設定)である。
以下にバイオセンサの製造方法について説明する。図12は、バイオセンサの製造方法を示す工程図である。
次に、図12のS2に示すように、試薬溶液が調整される。この試薬溶液としては、水に、少なくとも測定対象物を酸化又は脱水素する酸化還元酵素、酸化還元酵素と電子の授受を行うメディエータ及び光影響軽減剤を溶解させ、任意成分として、緩衝剤、高分子材料、酵素安定化剤、結晶均質化剤等を選択的に溶解させて作成される。
最後に、図12のS5に示すように、絶縁基板4、スペーサー10およびカバー11をこの順序で積層し、一体化し、バイオセンサが製造される。
上記実施の形態について、実施例を用いてより詳細に説明する。
(緑色3号の太陽光暴露影響)
図13(a)は、緑色3号を用いた際の太陽光暴露影響のグラフを示す図である。図13(b)は、青色1号を用いた際の太陽光暴露影響のグラフを示す図である。図13(c)は、黄色4号を用いた際の太陽光暴露影響のグラフを示す図である。
試薬溶液の組成は、wt(重量)%で水が91.25%、酵素としてグルコースデヒドロゲナーゼが7.41%、緑色3号(分子量808.87)が0.08%、緩衝剤としてK2HPO4、KH2PO4並びにNaH2PO4が全部で0.2%、接着剤として高分子材料であるCMC(カルボキシメチルセルロース)が0.13%、酵素安定化剤としてATA(3-アミノ-1,2,4-トリアゾール)が0.04%、保存安定化剤としてマルチトールが0.17%、メディエータとしてPQSAが0.70%、及びその他選択部材が0.03%となっている。これにより、緑色3号の濃度が1.0mMの試薬溶液が作成される。
これら色素なし試薬溶液、緑色3号1mMの試薬溶液、及び緑色3号10mMの試薬容器を用いてそれぞれ試薬部を作成し、その試薬部を用いて、太陽光に1時間暴露した場合と暴露していない場合においてサンプル溶液の測定結果の比較を行う。尚、図14(a)、(b)には、乾燥後の試薬部の各成分の重量(wt)%が示されている。
太陽光暴露なしの緑色3号1mM試薬を用いて、上記サンプル溶液を測定した場合の5回の平均値が41.7mg/dLであり、太陽光暴露有りの緑色3号1mM試薬を用いて上記サンプル溶液を測定した場合の5回の平均値が76.4mg/dLであった。このため、緑色3号0.1mM試薬を用いた場合の、光暴露を行っていない場合からの測定値の乖離は(76.4-41.7)/41.7=83.3であり、83.3%となる。
図13(b)では、例えば、図15(a)に示す表の試薬を水に添加して試薬溶液を作成し、その試薬溶液を乾燥させることによって、測定用試薬を作成した。
そして、緑色3号と同様の計算によってそれぞれの試薬部を用いた場合の乖離率を求めると、青色1号1mMの場合、乖離率は84.3%となり、青色1号5mMの場合、乖離率は24.7%となった。
図13(c)では、例えば、図16(a)に示す表の試薬を水に添加して溶液を作成し、その溶液を乾燥させることによって、測定用試薬を作成した。
(1)
上記実施の形態のバイオセンサ2は、血液成分測定作用極5と、血液成分測定対極6と、試薬部9とを備える。試薬部9は、血液成分測定作用極5と、血液成分測定対極6の近傍に設けられている。試薬部9は、メディエータと、酸化還元酵素と、光波長400nm~500nmを吸収する物質を含む。
(2)
上記実施の形態のバイオセンサ2は、血液成分測定作用極5と、血液成分測定対極6と、試薬部9とを備える。試薬部9は、血液成分測定作用極5と、血液成分測定対極6の近傍に設けられている。試薬部9は、メディエータと、酸化還元酵素と、光影響低減剤を含む。
これにより、メディエータが太陽光などに曝露されて劣化することを低減することが出来る。
また、上記実施の形態のバイオセンサ2は、血液成分測定作用極5及び血液成分測定対極6のうち少なくとも一方の電極が表面に配置された絶縁基板4(基板の一例)を備えている。試薬部9は、絶縁基板4の少なくとも一方の電極上に配置されている。
(4)
また、上記実施の形態のバイオセンサ2では、図5に示すように、試薬部9は、絶縁基板4上に配置された状態で、内側部分の肉厚d1よりも外周部分の肉厚d2が厚く形成されている。
また、上記実施の形態のバイオセンサ2では、図5に示すように、試薬部9は、絶縁基板4に配置された状態で、平面視において円形状である。
また、上記実施の形態のバイオセンサ2は、図3に示すように、スペーサー10と、カバー11とを備える。スペーサー10は、絶縁基板4の試薬部9が形成された側に積層されている。カバー11は、スペーサー10の絶縁基板4と反対側に積層されており、透明または半透明である。スペーサー10には、試薬部9に対応する部分に生体試料を試薬部9に導く生体試料導入部12が形成されている。
(7)
上記実施の形態において、光波長400nm~500nmを吸収する物質は、化学構造式において、疎水性ベンゼン環部分および親水性ベンゼン環部分を有する。
上記実施の形態において、光波長400nm~500nmを吸収する物質は、疎水性ベンゼン環部分を前記試薬部の表層側に向けて配置される。
(9)
上記実施の形態において、光波長400nm~500nmを吸収する物質は、ブリリアントブルーFCF(青色1号)ファストグリーンFCF(緑3号)もしくはタートラジン(黄色4号)である。
上記実施の形態において、試薬部9は、光波長400nm~500nmを吸収する物質のモル数は、メディエータのモル数よりも少ない。
(11)
上記実施の形態において、図13及び図14に示すようにファストグリーンFCFの試薬部9全体に対する重量割合は、6.01%以上、39.02%以下である。
(12)
上記実施の形態において、図13及び図14に示すようにファストグリーンFCFの試薬部9全体に対する重量割合は、メディエータの試薬部9全体に対する重量割合の11.6%以上、116%以下である。
(13)
上記実施の形態において、図13及び図15に示すようにブリリアントブルーFCFの試薬部9全体に対する重量割合は、5.9%以上、23.87%以下である。
上記実施の形態において、図13及び図15に示すようにブリリアントブルーFCFの試薬部9全体に対する重量割合は、メディエータの試薬部9全体に対する重量割合の11.4%以上、56.8%以下である。
上記実施の形態において、図13及び図16に示すようにタートラジンの試薬部9全体に対する重量割合は、4.06%以上、29.71%以下である。
上記実施の形態において、図13及び図16に示すようにタートラジンの試薬部9全体に対する重量割合は、メディエータの試薬部9全体に対する重量割合の7.66%以上、76.6%以下である。
上記実施の形態において、光影響軽減剤は、光波長400nm~500nmを吸収する物質である。
(18)
上記実施の形態において、バイオセンサの製造方法は、載置工程と、乾燥工程とを備えている。載置工程は、少なくとも酸化還元酵素、メディエータ及びファストグリーンFCFを含む試薬溶液を、基板上に形成された第1の電極及び第2の電極上に載置する。乾燥工程は、載置された前記試薬溶液を乾燥させる。試薬溶液におけるファストグリーンFCFの濃度は、1mM以上、10mM以下である。
(19)
上記実施の形態において、バイオセンサの製造方法は、載置工程と、乾燥工程とを備えている。載置工程は、少なくとも酸化還元酵素、メディエータ及びブリリアントブルーFCFを含む試薬溶液を、基板上に形成された第1の電極及び第2の電極上に載置する。乾燥工程は、載置された前記試薬溶液を乾燥させる。試薬溶液におけるブリリアントブルーFCFの濃度は、1mM以上、5mM以下である。
(20)
上記実施の形態において、バイオセンサの製造方法は、載置工程と、乾燥工程とを備える。載置工程は、少なくとも酸化還元酵素、メディエータ及びタートラジンを含む試薬溶液を、基板上に形成された第1の電極及び第2の電極上に載置する。乾燥工程は、載置された試薬溶液を乾燥させる。試薬溶液におけるタートラジンの濃度は、1mM以上、10mM以下である。
(他の実施の形態)
(A)
上記実施の形態では、メディエータの一例としてPQSAが用いられたが、これに限らず他のメディエータを用いてもよい。
また、本発明の試薬の組成は、上記実施の形態で用いた組成に限られない。例えば、上記実施の形態では、緩衝剤として、K2HPO40.0016g、KH2PO4並びにNaH2PO4が用いられているが、これらに限らず、HEPES等、他の緩衝剤が用いられても良い。
2 バイオセンサ
3 挿入口
4 絶縁基板(基板の一例)
5 血液成分測定作用極(第1の電極の一例)
6 血液成分測定対極(第2の電極の一例)
7 血液成分検知極
8 入力端子部
9 試薬部
10 スペーサー
11 カバー
12 生体試料導入路
13 血液供給口
14 空気孔
15 電圧印加部
16 電流-電圧変換部
17 切替回路
18 印加電圧部
19 基準電圧部
20 電源部
21 制御部
22 A/D変換部
23 判定手段
24 表示部
25 メモリ部
26 時計
27 補正手段
Claims (20)
- 第1の電極と、
第2の電極と、
前記第1の電極及び前記第2の電極の近傍に設けられた試薬部と、
を備え、
前記試薬部は、
メディエータと、
酸化還元酵素と、
光波長400nm~500nmを吸収する物質と、
を含む、バイオセンサ。 - 第1の電極と、
第2の電極と、
前記第1の電極及び前記第2の電極の近傍に設けられた試薬部と、
を備え、
前記試薬部は、
メディエータと、
酸化還元酵素と、
前記メディエータに対する光の影響を軽減する光影響軽減剤と、
を含む、バイオセンサ。 - 前記第1の電極及び前記第2の電極のうち少なくとも一方の電極が表面に配置された基板を備え、
前記試薬部は、前記基板上の前記少なくとも一方の電極上に配置された、
請求項1又は2に記載のバイオセンサ。 - 前記試薬部は、前記基板上に配置された状態で、内側部分の肉厚よりも外周部分の肉厚が厚く形成されている、
請求項3に記載のバイオセンサ。 - 前記試薬部は、前記基板上に配置された状態で、平面視において円形状である、
請求項4に記載のバイオセンサ。 - 前記基板の前記試薬部が形成された側に積層されたスペーサーと、
前記スペーサーの前記基板と反対側に積層された透明または半透明のカバーとを備え、
前記スペーサーには、前記試薬部に対応する部分に生体試料を前記試薬部に導く生体試料導入部が形成されている、
請求項3から5のいずれか一つに記載のバイオセンサ。 - 前記光波長400nm~500nmを吸収する物質は、化学構造式において、疎水性ベンゼン環部分および親水性ベンゼン環部分を有する、
請求項1、3から6のいずれか一つに記載のバイオセンサ。 - 前記光波長400nm~500nmを吸収する物質は、前記疎水性ベンゼン環部分を前記試薬部の表層側に向けて配置される、
請求項7に記載のバイオセンサ。 - 前記光波長400nm~500nmを吸収する物質は、ブリリアントブルーFCF、ファストグリーンFCFもしくはタートラジンである、
請求項1、3から8のいずれか一つに記載のバイオセンサ。 - 前記光波長400nm~500nmを吸収する物質の前記試薬部全体に対するモル数は、前記メディエータのモル数よりも少ない、
請求項1、3から9のいずれか一つに記載のバイオセンサ。 - 前記ファストグリーンFCFの前記試薬部全体に対する重量割合は、6.01%以上、39.02%以下である、
請求項1、3から10のいずれか1つに記載のバイオセンサ。 - 前記ファストグリーンFCFのモル数の前記メディエータのモル数に対する比は、0.0442以上、0.422以下である、請求項1及び3から10のいずれか1つに記載のバイオセンサ。
- 前記ブリリアントブルーFCFの前記試薬部全体に対する重量割合は、5.9%以上、23.87%以下である、
請求項1、3から10のいずれか1つに記載のバイオセンサ。 - 前記ブリリアントブルーFCFのモル数の前記メディエータのモル数に対する比は、0.0442以上、0.221以下である、請求項1、3から10のいずれか1つに記載のバイオセンサ。
- 前記タートラジンの前記試薬部全体に対する重量割合は、4.06%以上、29.71%以下である、
請求項1、3から10のいずれか1つに記載のバイオセンサ。 - 前記タートラジンのモル数の前記メディエータのモル数に対する比は、0.0422以上、0.442以下である、請求項1、3から10のいずれか1つに記載のバイオセンサ。
- 前記光影響軽減剤は、光波長400nm~500nmを吸収する物質である、請求項2記載のバイオセンサ。
- 少なくとも酸化還元酵素、メディエータ及びファストグリーンFCFを含む試薬溶液を、基板上に形成された第1の電極及び第2の電極上に載置する載置工程と、
載置された前記試薬溶液を乾燥させる乾燥工程とを備え、
前記試薬溶液における前記ファストグリーンFCFの濃度は、1mM以上、10mM以下である、
バイオセンサの製造方法。 - 少なくとも酸化還元酵素、メディエータ及びブリリアントブルーFCFを含む試薬溶液を、基板上に形成された第1の電極及び第2の電極上に載置する載置工程と、
載置された前記試薬溶液を乾燥させる乾燥工程とを備え、
前記試薬溶液における前記ブリリアントブルーFCFの濃度は、1mM以上、5mM以下である、
バイオセンサの製造方法。 - 少なくとも酸化還元酵素、メディエータ及びタートラジンを含む試薬溶液を、基板上に形成された第1の電極及び第2の電極上に載置する載置工程と、
載置された前記試薬溶液を乾燥させる乾燥工程とを備え、
前記試薬溶液における前記タートラジンの濃度は、1mM以上、10mM以下である、
バイオセンサの製造方法。
Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP13839044.8A EP2899538B1 (en) | 2012-09-19 | 2013-09-19 | Biosensor |
JP2014536601A JP6282226B2 (ja) | 2012-09-19 | 2013-09-19 | バイオセンサ及びバイオセンサの製造方法 |
CN201380048522.2A CN104641227B (zh) | 2012-09-19 | 2013-09-19 | 生物传感器及生物传感器的制造方法 |
US14/426,642 US9588076B2 (en) | 2012-09-19 | 2013-09-19 | Biosensor and method for manufacturing biosensor |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2012205274 | 2012-09-19 | ||
JP2012-205274 | 2012-09-19 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2014045584A1 true WO2014045584A1 (ja) | 2014-03-27 |
Family
ID=50340920
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/JP2013/005542 WO2014045584A1 (ja) | 2012-09-19 | 2013-09-19 | バイオセンサ及びバイオセンサの製造方法 |
Country Status (5)
Country | Link |
---|---|
US (1) | US9588076B2 (ja) |
EP (1) | EP2899538B1 (ja) |
JP (1) | JP6282226B2 (ja) |
CN (1) | CN104641227B (ja) |
WO (1) | WO2014045584A1 (ja) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2019526049A (ja) * | 2016-07-19 | 2019-09-12 | バイオメトリー・インコーポレイテッドBiometry Inc. | バッチ校正可能なテストストリップを用いる検体測定方法と検体測定システム |
US11175268B2 (en) | 2014-06-09 | 2021-11-16 | Biometry Inc. | Mini point of care gas chromatographic test strip and method to measure analytes |
US11435340B2 (en) | 2014-06-09 | 2022-09-06 | Biometry Inc. | Low cost test strip and method to measure analyte |
Families Citing this family (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20140152801A1 (en) | 2009-10-28 | 2014-06-05 | Alentic Microscience Inc. | Detecting and Using Light Representative of a Sample |
US10502666B2 (en) * | 2013-02-06 | 2019-12-10 | Alentic Microscience Inc. | Sample processing improvements for quantitative microscopy |
EP3014330B1 (en) | 2013-06-26 | 2024-01-03 | Alentic Microscience Inc. | Sample processing improvements for microscopy |
KR102197061B1 (ko) * | 2017-10-23 | 2020-12-30 | 주식회사 비바이오 | 커피링 효과에 강인한 바이오센서 |
JPWO2019187575A1 (ja) * | 2018-03-26 | 2021-03-11 | Phcホールディングス株式会社 | 生体物質検出用センサ |
EP3771411A1 (en) * | 2019-08-02 | 2021-02-03 | Bionime Corporation | Method for reducing measurement interference of micro biosensor |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2004092720A1 (ja) | 2003-03-25 | 2004-10-28 | Arkray Inc. | センサ収納容器 |
JP2006170631A (ja) * | 2004-12-13 | 2006-06-29 | Matsushita Electric Ind Co Ltd | バイオセンサ、及びその検査装置、ならびにその検査方法 |
JP2009210347A (ja) * | 2008-03-03 | 2009-09-17 | Nokodai Tlo Kk | インジケータ |
JP2012504233A (ja) * | 2008-09-30 | 2012-02-16 | メナイ メディカル テクノロジーズ リミテッド | サンプル測定システム |
Family Cites Families (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5126247A (en) * | 1988-02-26 | 1992-06-30 | Enzymatics, Inc. | Method, system and devices for the assay and detection of biochemical molecules |
MXPA05013635A (es) | 2003-06-20 | 2006-02-24 | Hoffmann La Roche | Metodo y reactivo para producir tiras reactivas estrechas y homogeneas. |
US20070235346A1 (en) * | 2006-04-11 | 2007-10-11 | Popovich Natasha D | System and methods for providing corrected analyte concentration measurements |
WO2008007277A2 (en) * | 2006-06-27 | 2008-01-17 | Agamatrix, Inc. | Detection of analytes in a dual-mediator electrochemical test strip |
US8262874B2 (en) * | 2008-04-14 | 2012-09-11 | Abbott Diabetes Care Inc. | Biosensor coating composition and methods thereof |
JP5091885B2 (ja) | 2009-02-12 | 2012-12-05 | 株式会社リコー | 定着装置及び画像形成装置 |
US20110046466A1 (en) * | 2009-08-19 | 2011-02-24 | Feldman Benjamin J | Analyte Sensors Including Nanomaterials and Methods of Using Same |
EP4397242A2 (en) * | 2010-12-09 | 2024-07-10 | Abbott Diabetes Care Inc. | Analyte sensors with a sensing surface having small sensing spots |
-
2013
- 2013-09-19 WO PCT/JP2013/005542 patent/WO2014045584A1/ja active Application Filing
- 2013-09-19 CN CN201380048522.2A patent/CN104641227B/zh active Active
- 2013-09-19 US US14/426,642 patent/US9588076B2/en active Active
- 2013-09-19 JP JP2014536601A patent/JP6282226B2/ja active Active
- 2013-09-19 EP EP13839044.8A patent/EP2899538B1/en active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2004092720A1 (ja) | 2003-03-25 | 2004-10-28 | Arkray Inc. | センサ収納容器 |
JP2006170631A (ja) * | 2004-12-13 | 2006-06-29 | Matsushita Electric Ind Co Ltd | バイオセンサ、及びその検査装置、ならびにその検査方法 |
JP2009210347A (ja) * | 2008-03-03 | 2009-09-17 | Nokodai Tlo Kk | インジケータ |
JP2012504233A (ja) * | 2008-09-30 | 2012-02-16 | メナイ メディカル テクノロジーズ リミテッド | サンプル測定システム |
Non-Patent Citations (1)
Title |
---|
See also references of EP2899538A4 |
Cited By (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US11175268B2 (en) | 2014-06-09 | 2021-11-16 | Biometry Inc. | Mini point of care gas chromatographic test strip and method to measure analytes |
US11435340B2 (en) | 2014-06-09 | 2022-09-06 | Biometry Inc. | Low cost test strip and method to measure analyte |
US11747324B2 (en) | 2014-06-09 | 2023-09-05 | Biometry Inc. | Low cost test strip and method to measure analyte |
JP2019526049A (ja) * | 2016-07-19 | 2019-09-12 | バイオメトリー・インコーポレイテッドBiometry Inc. | バッチ校正可能なテストストリップを用いる検体測定方法と検体測定システム |
US11255840B2 (en) | 2016-07-19 | 2022-02-22 | Biometry Inc. | Methods of and systems for measuring analytes using batch calibratable test strips |
JP7086927B2 (ja) | 2016-07-19 | 2022-06-20 | バイオメトリー・インコーポレイテッド | バッチ校正可能なテストストリップを用いる検体測定方法と検体測定システム |
JP2022120051A (ja) * | 2016-07-19 | 2022-08-17 | バイオメトリー・インコーポレイテッド | バッチ校正可能なテストストリップを用いる検体測定方法と検体測定システム |
JP7358563B2 (ja) | 2016-07-19 | 2023-10-10 | バイオメトリー・インコーポレイテッド | バッチ校正可能なテストストリップを用いる検体測定方法と検体測定システム |
US12038428B2 (en) | 2016-07-19 | 2024-07-16 | Biometry Inc. | Methods of and systems for measuring analytes using batch calibratable test strips |
Also Published As
Publication number | Publication date |
---|---|
JPWO2014045584A1 (ja) | 2016-08-18 |
CN104641227A (zh) | 2015-05-20 |
EP2899538B1 (en) | 2016-11-09 |
CN104641227B (zh) | 2016-10-05 |
JP6282226B2 (ja) | 2018-02-21 |
EP2899538A4 (en) | 2015-10-21 |
US9588076B2 (en) | 2017-03-07 |
US20150241377A1 (en) | 2015-08-27 |
EP2899538A1 (en) | 2015-07-29 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP6282226B2 (ja) | バイオセンサ及びバイオセンサの製造方法 | |
EP1909097B1 (en) | Biosensor and biosensor cell | |
AU2016203268B2 (en) | Reagent composition for biosensor and biosensor having the same | |
JP6338748B2 (ja) | センサ | |
US20130098775A1 (en) | Glucose biosensor with improved shelf life | |
EP3292398B1 (en) | Electrochemical test device | |
JP5940719B2 (ja) | 生体情報測定装置とそれを用いた生体情報測定方法 | |
JP2000065778A (ja) | バイオセンサ | |
WO2002008743A1 (fr) | Biocapteur | |
JP2014006155A (ja) | 酵素電極 | |
US10371617B2 (en) | Electrochemical test device | |
JP2002207022A (ja) | バイオセンサ | |
RU2263904C2 (ru) | Сенсор на антиоксиданты | |
JPH11344461A (ja) | バイオセンサ | |
US10775331B2 (en) | Biological information measuring device, bio sensor system, and error detection method for biological information measuring device | |
WO2015002184A1 (ja) | バイオセンサ | |
US10627361B2 (en) | Electrochemical test device | |
JP7362890B2 (ja) | センサーおよびその製造方法 | |
FR2956904A1 (fr) | Procede de determination de substances oxydantes et reductrices dans un aliment, echantillon d'analyse et dispositif de mesure pour ce procede | |
JP2005010150A (ja) | バイオセンサ |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 13839044 Country of ref document: EP Kind code of ref document: A1 |
|
REEP | Request for entry into the european phase |
Ref document number: 2013839044 Country of ref document: EP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2013839044 Country of ref document: EP |
|
ENP | Entry into the national phase |
Ref document number: 2014536601 Country of ref document: JP Kind code of ref document: A |
|
WWE | Wipo information: entry into national phase |
Ref document number: 14426642 Country of ref document: US |
|
NENP | Non-entry into the national phase |
Ref country code: DE |