US20050133368A1 - Electrochemical test strip for reducing the effect of direct interference current - Google Patents

Electrochemical test strip for reducing the effect of direct interference current Download PDF

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Publication number
US20050133368A1
US20050133368A1 US10/976,489 US97648904A US2005133368A1 US 20050133368 A1 US20050133368 A1 US 20050133368A1 US 97648904 A US97648904 A US 97648904A US 2005133368 A1 US2005133368 A1 US 2005133368A1
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working electrode
electrode
layer
test strip
working
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US10/976,489
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Oliver Davies
Robert Marshall
Damian Baskeyfield
Lynsey Whyte
Elaine Leiper
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LifeScan Scotland Ltd
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Individual
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Assigned to LIFESCAN, SCOTLAND, LTD. reassignment LIFESCAN, SCOTLAND, LTD. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: BASKEYFIELD, DAMIAN EDWARD HAYDON, DAVIES, OLIVER WILLIAM HARDWICKE, LEIPER, ELAINE, MARSHALL, ROBERT, WHYTE, LYNSEY
Publication of US20050133368A1 publication Critical patent/US20050133368A1/en
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    • 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
    • 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
    • G01N27/416—Systems
    • G01N27/49—Systems involving the determination of the current at a single specific value, or small range of values, of applied voltage for producing selective measurement of one or more particular ionic species
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/145—Measuring characteristics of blood in vivo, e.g. gas concentration or pH-value ; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid or cerebral tissue
    • A61B5/14532—Measuring characteristics of blood in vivo, e.g. gas concentration or pH-value ; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid or cerebral tissue for measuring glucose, e.g. by tissue impedance measurement
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/145—Measuring characteristics of blood in vivo, e.g. gas concentration or pH-value ; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid or cerebral tissue
    • A61B5/1468—Measuring characteristics of blood in vivo, e.g. gas concentration or pH-value ; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid or cerebral tissue using chemical or electrochemical methods, e.g. by polarographic means
    • A61B5/1486—Measuring characteristics of blood in vivo, e.g. gas concentration or pH-value ; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid or cerebral tissue using chemical or electrochemical methods, e.g. by polarographic means using enzyme electrodes, e.g. with immobilised oxidase
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/15—Devices for taking samples of blood
    • A61B5/150007—Details
    • A61B5/150015—Source of blood
    • A61B5/150022—Source of blood for capillary blood or interstitial fluid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/15—Devices for taking samples of blood
    • A61B5/150007—Details
    • A61B5/150206—Construction or design features not otherwise provided for; manufacturing or production; packages; sterilisation of piercing element, piercing device or sampling device
    • A61B5/150274—Manufacture or production processes or steps for blood sampling devices
    • A61B5/150282—Manufacture or production processes or steps for blood sampling devices for piercing elements, e.g. blade, lancet, canula, needle
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/15—Devices for taking samples of blood
    • A61B5/150007—Details
    • A61B5/150358—Strips for collecting blood, e.g. absorbent
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/15—Devices for taking samples of blood
    • A61B5/150007—Details
    • A61B5/150374—Details of piercing elements or protective means for preventing accidental injuries by such piercing elements
    • A61B5/150381—Design of piercing elements
    • A61B5/150412—Pointed piercing elements, e.g. needles, lancets for piercing the skin
    • A61B5/150435—Specific design of proximal end
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/15—Devices for taking samples of blood
    • A61B5/150007—Details
    • A61B5/150374—Details of piercing elements or protective means for preventing accidental injuries by such piercing elements
    • A61B5/150381—Design of piercing elements
    • A61B5/150503—Single-ended needles
    • 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
    • G01N27/3274—Corrective measures, e.g. error detection, compensation for temperature or hematocrit, calibration
    • 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
    • G01N27/416—Systems
    • 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
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A90/00—Technologies having an indirect contribution to adaptation to climate change
    • Y02A90/10—Information and communication technologies [ICT] supporting adaptation to climate change, e.g. for weather forecasting or climate simulation

Definitions

  • the present invention is related to the following co-pending U.S. applications: U.S. patent application Ser. No. ______ [Attorney Docket Number DDI-5027 USNP], filed on Oct. 29, 2004; U.S. patent application Ser. No. ______ [Attorney Docket Number DDI-5042 USNP], filed on Oct. 29, 2004; U.S. patent application Ser. No. ______ [Attorney Docket Number DDI-5065], filed on Oct. 29, 2004; U.S. patent application Ser. No. ______ [Attorney Docket Number DDI-5066], filed on Oct. 29, 2004; and U.S. patent application Ser. No. ______ [Attorney Docket Number DDI-5067], filed on Oct. 29, 2004.
  • the present invention is related, in general to electrochemical strips and systems which are designed to reduce the effect of interfering compounds on measurements taken by such analyte measurement systems and, more particularly, to an improved electrochemical strip for reducing the effects of direct interference currents in a glucose monitoring system wherein the electrochemical strip has electrodes with uncoated regions.
  • an electrochemical glucose measuring system may have an elevated oxidation current due to the oxidation of interfering compounds commonly found in physiological fluids such as, for example, acetaminophen, ascorbic acid, bilirubin, dopamine, gentisic acid, glutathione, levodopa, methyldopa, tolazimide, tolbutamide, and uric acid.
  • the accuracy of glucose meters may, therefore, be improved by reducing or eliminating the portion of the oxidation current generated by interfering compounds.
  • there should be no oxidation current generated from any of the interfering compounds so that the entire oxidation current would depend only on the glucose concentration.
  • electrochemical sensors it is, therefore, desirable to improve the accuracy of electrochemical sensors in the presence of potentially interfering compounds such as, for example, ascorbate, urate, and, acetaminophen, commonly found in physiological fluids.
  • analytes for such electrochemical sensors may include glucose, lactate, and fructosamine.
  • glucose will be the main analyte discussed, it will be obvious to one skilled in the art that the invention set forth herein may also be used with other analytes.
  • Oxidation current may be generated in several ways.
  • desirable oxidation current results from the interaction of the mediator with the analyte of interest (e.g., glucose) while undesirable oxidation current is generally comprised of interfering compounds being oxidized at the electrode surface and by interaction with the mediator.
  • interfering compounds e.g., acetominophen
  • Other interfering compounds e.g., ascorbic acid
  • This oxidation of the interfering compound in a glucose measuring system causes the measured oxidation current to be dependent on the concentration of both the glucose and any interfering compound.
  • the measurement of the glucose concentration would be improved by reducing or eliminating the contribution of the interfering compounds to the total oxidation current.
  • a negatively charged membrane to cover the working electrode.
  • a sulfonated fluoropolymer such as NAFIONTM may be used to repel all negatively charged chemicals.
  • most interfering compounds such as ascorbate and urate have a negative charge, thus, the negatively charged membrane prevents the negatively charged interfering compounds from reaching the electrode surface and being oxidized at that surface.
  • this technique is not always successful since some interfering compounds such as acetaminophen do not have a net negative charge, and thus, can pass through a negatively charged membrane.
  • This technique reduce the oxidation current resulting from the interaction of interfering compounds with some mediators.
  • the use of a negatively charged membrane on the working electrode could also prevent some commonly used mediators, such as ferricyanide, from passing through the negatively charged membrane to exchange electrons with the electrode.
  • a size selective membrane on top of the working electrode.
  • a 100 Dalton exclusion membrane such as cellulose acetate may be used to cover the working electrode to exclude all chemicals with a molecular weight greater than 100 Daltons.
  • most interfering compounds have a molecular weight greater than 100 Daltons, and thus, are excluded from being oxidized at the electrode surface.
  • selective membranes typically make the test strip more complicated to manufacture and increase the test time because the oxidized glucose must diffuse through the selective membrane to get to the electrode.
  • mediators having a relatively low redox potential include osmium bipyridyl complexes, ferrocene derivatives, and quinone derivatives.
  • mediators having a relatively low potential are often difficult to synthesize, unstable and have a low water solubility.
  • Another known strategy that can be used to decrease the effects of interfering compounds is to use a dummy electrode which is coated with a mediator.
  • the dummy electrode may also be coated with an inert protein or deactivated redox enzyme.
  • the purpose of the dummy electrode is to oxidize the interfering compound at the electrode surface and/or to oxidize the mediator reduced by the interfering compound.
  • the current measured at the dummy electrode is subtracted from the total oxidizing current measured at the working electrode to remove the interference effect.
  • a disadvantage of this strategy is that it requires that the test strip include an additional electrode and electrical connection (i.e., the dummy electrode) which cannot be used to measure glucose.
  • the inclusion of dummy electrode is an inefficient use of an electrode in a glucose measuring system.
  • An electrochemical sensor according to the present invention includes a substrate, at least first and second working electrodes and a reference electrode.
  • a reagent layer is disposed on the electrodes such that it completely covers all of the first working electrode and only partially covers the second working electrode.
  • the oxidation current generated at the portion of the second working electrode not covered by the reagent layer is used to correct for the effect of interfering substances on the glucose measurement.
  • the electrochemical glucose test strip includes a first and second working electrodes, where the first working electrode is completely covered with a reagent layer and the second working electrode is only partially covered with the reagent layer.
  • the second working electrode has a reagent coated area and an uncoated area.
  • the reagent layer may include, for example, a redox enzyme such as glucose oxidase and a mediator such as, for example, ferricyanide.
  • the first working electrode will have a superposition of two oxidation current sources, one from glucose and a second from interferents.
  • the second working electrode will have a superposition of three oxidation current sources from glucose, interferents at the reagent coated portion, and interferents at the uncoated portion.
  • the uncoated portion of the second working electrode will only oxidize interferents and not oxidize glucose because there is no reagent is in this area.
  • the oxidation current measured at the uncoated portion of the second working electrode may then be used to estimate the total interferent oxidation current and calculate a corrected oxidation current which removes the effects of interferences.
  • the electrochemical glucose test strip includes a first and second working electrodes, where the first and second working electrode are only partially covered with the reagent layer.
  • both the first and second working electrode have a reagent coated portion and an uncoated portion.
  • the first uncovered area of the first working electrode and the second uncovered area of the second working electrode are different.
  • the oxidation current measured at the uncoated portion of the first and second working electrodes are used to estimate the interferent oxidation current for the uncoated portion and to calculate a corrected glucose current.
  • FIG. 1 is an exploded perspective view of a test strip according to an embodiment of the present invention
  • FIG. 2 is a simplified plane view of a distal portion of a test strip according to the embodiment of the present invention illustrated in FIG. 1 including a conductive layer and an insulation layer;
  • FIG. 3 is a simplified plane view of a distal portion of a test strip according to the embodiment of the present invention illustrated in FIG. 1 wherein the position of a reagent layer is illustrated with the conductive layer and the insulation layer;
  • FIG. 4 is an exploded perspective view of a test strip according to a further embodiment of the present invention.
  • FIG. 5 is a simplified plane view of a distal portion of a test strip according to the embodiment of the present invention illustrated in FIG. 4 including of a conductive layer and an insulation layer;
  • FIG. 6 is a simplified plane view of a distal portion of a test strip according to the embodiment of the present invention illustrated in FIG. 4 wherein a reagent layer is illustrated with the conductive layer and the insulation layer.
  • FIG. 7 is a simplified plane view of a distal portion of a test strip according to the embodiment of the present invention illustrated in FIG. 4 wherein a reagent layer is illustrated with the conductive layer.
  • FIG. 8 is a simplified plane view of a distal portion of a test strip according to another embodiment of the present invention wherein a reagent layer is illustrated with the conductive layer that helps reduce an IR drop effect.
  • FIG. 9 is a simplified plane view of a distal portion of a test strip according to yet another embodiment of the present invention wherein a reagent layer is illustrated with the conductive layer and the insulation layer such there are two working electrodes that have an uncoated portion.
  • FIG. 10 is a simplified plane view of a distal portion of a test strip according to still yet another embodiment of the present invention wherein a reagent layer is illustrated with the conductive layer and the insulation layer such there are two working electrodes that have an uncoated portion.
  • FIG. 11 is a graph showing the current at a first working electrode of a strip designed in accordance with the present invention tested with 70 mg/dL glucose samples in blood spiked with varying levels of uric acid.
  • FIG. 12 is a graph showing the current at a first working electrode at a strip designed in accordance with the present invention tested with 240 mg/dL glucose samples in blood spiked with varying levels of uric acid.
  • FIG. 13 is an exploded perspective view of a test strip that has an integrated lance.
  • This invention described herein includes a test strip and method for improving the selectivity of an electrochemical glucose measuring system.
  • FIG. 1 is an exploded perspective view of a test strip according to a first embodiment of the present invention.
  • an electrochemical test strip 62 which may be used for measuring glucose concentration in bodily fluids such as blood or interstitial fluid, includes a first working electrode 10 and a second working electrode 12 , where first working electrode 10 is completely covered with a reagent layer 22 and second working electrode 12 is only partially covered with reagent layer 22 .
  • the second working electrode has a reagent coated portion and an uncoated portion.
  • Reagent layer 22 may include, for example, a redox enzyme such as, for example, glucose oxidase and a mediator such as, for example, ferricyanide.
  • ferricyanide has a redox potential of approximately 400 mV (when measured with respect to a saturated calomel electrode) at a carbon electrode
  • the introduction of a bodily fluid e.g., blood may generate a significant oxidation of interferents by the mediator and/or the working electrode. Therefore, the oxidation current measured at first working electrode 10 will be a superposition of oxidation current sources: a first, desirable, oxidation current generated by the oxidation of glucose and a second, undesirable, oxidation current generated by the interferents.
  • the oxidation current measured at second working electrode 12 will also be a superposition of oxidation current sources: a first, desirable oxidation current generated by the oxidation of glucose, a second, undesirable oxidation current generated by interferents at the covered portion of working electrode 12 and a third oxidation current generated by interferents at the uncovered portion of working electrode 12 .
  • the uncoated portion of second working electrode 12 will only oxidize interferents and not oxidize glucose because there is no reagent on the uncoated portion of second working electrode 12 .
  • the oxidation current measured at the uncoated portion of second working electrode 12 does not depend on glucose and the uncoated area of second working electrode 12 is known, it is possible to calculate the interferent oxidation current for the uncoated portion of the second working electrode 12 . In turn, using the interferent oxidation current calculated for the uncoated portion of second working electrode 12 and knowing the area of first working electrode 10 and the area of the coated portion of second working electrode 12 , it is possible to calculate a corrected glucose current which accounts for the effects of interfering compounds oxidized at the electrode.
  • FIG. 1 is an exploded perspective view of a test strip 62 according to a first embodiment of the present invention.
  • Test strip 62 may be manufactured by a series of 6 consecutive printing steps which lay down six layers of material on substrate 50 .
  • the six layers may be deposited by, for example, screen printing on substrate 50 .
  • the 6 layers may include a conductive layer 64 , an insulation layer 16 , a reagent layer 22 , an adhesive layer 66 , a hydrophilic layer 68 , and a top layer 40 .
  • Conductive layer 64 may further includes first working electrode 10 , second working electrode 12 , reference electrode 14 , first contact 11 , second contact 13 , reference contact 15 , and strip detection bar 17 .
  • Insulation layer 16 may further include cutout 18 .
  • Adhesive layer 66 may further include first adhesive pad 24 , second adhesive pad 26 , and third adhesive pads 28 .
  • Hydrophilic layer 68 may further include first hydrophilic film 32 , and second hydrophilic film 34 .
  • Top layer 40 may further includes a clear portion 36 and opaque portion 38 .
  • Test strip 62 has a first side 54 and second side 56 , a distal electrode side 58 , and a proximal electrode side 60 as illustrated in FIG. 1 . The following sections will describe the respective layers of test strip 62 in more detail.
  • substrate 50 is an electrically insulating material such as plastic, glass, ceramic, and the like.
  • substrate 50 may be a plastic such as, for example, nylon, polycarbonate, polyimide, polyvinylchloride, polyethylene, polypropylene, PETG, or polyester. More particularly the polyester may be, for example Melinex® ST328 which is manufactured by DuPont Teijin Films.
  • Substrate 50 may also include an acrylic coating which is applied to one or both sides to improve ink adhesion.
  • the first layer deposited on substrate 50 is conductive layer 64 which includes first working electrode 10 , second working electrode 12 , reference electrode 14 , and strip detection bar 17 .
  • a screen mesh with an emulsion pattern may be used to deposit a material such as, for example, a conductive carbon ink in a defined geometry as illustrated in FIG. 1 .
  • Reference electrode 14 may also be a counter electrode, a reference/counter electrode, or a quasi-reference electrode.
  • Conductive layer 64 may be disposed on substrate 50 by using screen printing, rotogravure printing, sputtering, evaporation, electroless plating, ink jetting, sublimation, chemical vapor deposition, and the like.
  • Suitable materials which may be used for conductive layer 64 are Au, Pd, Ir, Pt, Rh, stainless steel, doped tin oxide, carbon, and the like.
  • the carbon ink layer may have a height between 1 and 100 microns, more particularly between 5 and 25 microns, and yet even more particularly at approximately 13 microns.
  • the height of the conductive layer can vary depending on the desired resistance of the conductive layer and the conductivity of the material used for printing the conductive layer.
  • First contact 11 , second contact 13 , and reference contact 15 may be used to electrically interface with a meter. This allows the meter to electrically communicate to first working electrode 10 , second working electrode 12 , and reference electrode 14 via, respective, first contact 11 , second contact 13 , and reference contact 15 .
  • the second layer deposited on substrate 50 is insulation layer 16 .
  • Insulation layer 16 is disposed on at least a portion of conductive layer 64 as shown in FIG. 1 .
  • FIG. 2 is a simplified plane view of a distal portion of test strip 62 which highlights the position of first working electrode 10 , second working electrode 12 , and reference electrode 14 with respect to insulation layer 16 .
  • Insulation layer 16 further includes a cutout 18 which may have a T-shaped structure as shown in FIGS. 1 and 2 . Cutout 18 exposes a portions of first working electrode 10 , second working electrode 12 , and reference electrode 14 which can be wetted with liquid.
  • distal cutout width W 1 , proximal cutout width W 2 , distal cutout length L 4 and proximal cutout length L 5 may have a respective dimension of approximately 0.7, 1.9, 3.2, and 0.43 mm.
  • first working electrode 10 , reference electrode 14 , and second working electrode 12 have a respective length of L 1 , L 2 , and L 3 which may be about 0.8, 1.6, and 0.4 mm.
  • electrode spacing S 1 is a distance between first working electrode 10 and reference electrode 14 ; and between reference electrode 14 and second working electrode 12 which may be about 0.4 mm.
  • the third layer deposited on substrate 50 is a reagent layer 22 .
  • Reagent layer 22 is disposed on at least a portion of conductive layer 64 and insulation layer 16 as shown in FIGS. 1 .
  • FIG. 3 is a simplified plane view of a distal portion of test strip 62 according to the first embodiment of the present invention which highlights the position of reagent layer 22 with respect to first working electrode 10 , second working electrode 12 , reference electrode 14 , and insulation layer 16 .
  • Reagent layer 22 may be in the shape of a rectangle having a reagent width W 3 and a reagent length L 6 as illustrated in FIGS. 1 and 3 .
  • reagent width W 3 may be about 1.3 mm and reagent length L 6 may be about 4.7 mm.
  • reagent layer 22 has a sufficiently large width W 3 and length L 6 such that reagent layer 22 completely covers first working electrode 10 and reference electrode 14 .
  • reagent layer 22 has an appropriately sized width W 3 and length L 6 such that second working electrode is not completely covered with reagent layer 22 .
  • second working electrode 12 has a coated portion 12 c and an uncoated portions 12 u as illustrated in FIG. 3 .
  • Uncoated portions 12 u may be in the shape of two rectangles where uncoated portions 12 u has a wing width W 4 and a length that corresponds to second working electrode length L 3 .
  • wing width W 4 may be about 0.3 mm.
  • reagent layer 22 may include a redox enzyme such as, for example, glucose oxidase or PQQ-glucose dehydrogenase (where PQQ is an acronym for pyrrolo-quinoline-quinone) and a mediator such as, for example, ferricyanide.
  • the fourth layer deposited on substrate 50 is an adhesive layer 66 which includes a first adhesive pad 24 , a second adhesive pad 26 , and a third adhesive pad 28 .
  • First adhesive pad 24 and second adhesive pad 26 form the walls of a sample receiving chamber.
  • first adhesive pad 24 and second adhesive pad 26 may be disposed on substrate 50 such that neither of the adhesive pads touches reagent layer 22 .
  • first adhesive pad 24 and/or second adhesive pad 26 may be disposed on substrate 50 such there is overlap with reagent layer 22 .
  • adhesive layer 66 has a height of about 70 to 110 microns.
  • Adhesive layer 66 may include a double sided pressure sensitive adhesive, a UV cured adhesive, heat activated adhesive, thermosetting plastic, or other adhesive known to those skilled in the art.
  • adhesive layer 66 may be formed by screen printing a pressure sensitive adhesive such as, for example, a water based acrylic copolymer pressure sensitive adhesive which is commercially available from Tape Specialties LTD in Tring, Herts, United Kingdom (part#A6435).
  • the fifth layer deposited on substrate 50 is a hydrophilic layer 68 which includes a first hydrophilic film 32 and second hydrophilic film 34 as illustrated in FIG. 1 .
  • Hydrophilic layer 68 forms the “roof” of the sample receiving chamber.
  • the “side walls” and “floor” of the sample receiving chamber are formed by a portion of adhesive layer 66 and substrate 50 , respectively.
  • hydrophilic layer 68 may be an optically transparent polyester with a hydrophilic anti-fog coating such as those commercially obtained from 3M. The hydrophilic nature of the coating is used in the design of strip 62 because it facilitates filling of liquid into the sample receiving chamber.
  • top layer 40 which includes a clear portion 36 and opaque portion 38 as illustrated in FIG. 1 .
  • top layer 40 includes a polyester which is coated on one side with a pressure sensitive adhesive.
  • Top layer 40 has an opaque portion 38 which helps the user observe a high degree of contrast when blood is underneath clear portion 36 . This allows a user to visually confirm that the sample receiving chamber is sufficiently filled. After strip 62 is fully laminated, it is cut along incision line A-A′ and in the process creates sample inlet 52 as illustrated in FIG. 3 .
  • the first test strip embodiment as illustrated in FIGS. 1-3 may have a possible drawback in that reagent layer 22 may dissolve in a liquid sample and move a portion of the dissolved reagent layer over the uncoated portions 12 u of second working electrode 12 . If such a scenario were to occur, uncoated portions 12 u would also measure an oxidation current that is also proportional to the glucose concentration. This would degrade the ability to use mathematical algorithms for removing the effect of interferent oxidation.
  • reagent layer 22 should be designed to dissolve in such a way that it does not migrate to uncoated portions 12 u .
  • reagent layer 22 may be chemically bound to the first working electrode 10 , second working electrode 12 , and reference electrode 14 or may have a thickening agent that minimizes the migration of dissolved reagent layer 22 .
  • second working electrode 102 has a C-shaped geometry where 2 discrete portions of second working electrode 102 are exposed by cutout 108 as illustrated in FIG. 4 .
  • reagent layer 110 is disposed on only a portion of second working electrode 102 to form an uncoated portion 102 u and coated portion 102 c as illustrated in FIG. 6 .
  • Uncoated portion 102 u is adjacent to sample inlet 52 .
  • Coated portion 102 c is adjacent to first working electrode 100 .
  • FIG. 4 is an exploded perspective view of a test strip 162 .
  • Test strip 162 is manufactured in a manner similar to test strip 62 except that there are geometric or positional changes to a conductive layer 164 , an insulation layer 106 , and a reagent layer 110 .
  • substrate 50 , adhesive layer 66 , hydrophilic layer 68 , and top layer 40 are the same as the first strip embodiment.
  • Test strip 162 has a first side 54 and second side 56 , a distal electrode side 58 , and a proximal electrode side 60 .
  • the first and second test strip embodiment of the present invention may have elements with similar structure which are denoted with the same element number and name. If analogous elements between the respective test strip embodiments are different in structure, the elements may have the same name, but be denoted with a different element number. The following sections will describe the respective layers of test strip 162 in more detail.
  • the first layer deposited on substrate 50 is conductive layer 164 which includes first working electrode 100 , second working electrode 102 , reference electrode 104 , first contact 101 , second contact 103 , and reference contact 105 , and strip detection bar 17 .
  • a screen mesh with an emulsion pattern may be used to deposit a material such as, for example, a conductive carbon ink in a defined geometry as illustrated in FIG. 4 .
  • First contact 101 , second contact 103 , and reference contact 105 may be used to electrically interface with a meter. This allows the meter to electrically communicate to first working electrode 100 , second working electrode 102 , and reference electrode 104 via, respective, first contact 101 , second contact 103 , and reference contact 105 .
  • the second layer deposited on substrate 50 in FIG. 4 is insulation layer 106 .
  • Insulation layer 106 is disposed on at least a portion of conductive layer 164 as shown in FIG. 4 .
  • FIG. 5 is a simplified plane view of a distal portion of test strip 162 which highlights the position of first working electrode 100 , second working electrode 102 , and reference electrode 104 with respect to insulation layer 106 .
  • the third layer deposited on substrate 50 in FIG. 4 is a reagent layer 110 such that reagent layer 110 is disposed on at least a portion of conductive layer 164 and insulation layer 106 as shown in FIG. 6 .
  • FIG. 6 is a simplified plane view of a distal portion of test strip 162 according to the second embodiment of the present invention which highlights the position of reagent layer 110 with respect to first working electrode 100 , second working electrode 102 , reference electrode 104 , and insulation layer 106 .
  • Reagent layer 110 may be in the shape of a rectangle having a reagent width W 13 and a reagent length L 16 . In one embodiment of this invention, reagent width W 13 may be about 1.3 mm and reagent length L 16 may be about 3.2 mm.
  • reagent layer 110 has a sufficient width W 13 and length L 16 such that reagent layer 110 completely covers first working electrode 100 , coated portion 102 c , and reference electrode 104 , but does not cover uncoated portion 102 u.
  • FIG. 7 is a simplified plane view of a distal portion of a test strip according to the embodiment of the present invention illustrated in FIG. 4 wherein a reagent layer is illustrated with the conductive layer. In contrast to FIG. 6 , FIG. 7 does not show insulation layer 106 . This helps demonstrate the conductive relationship between uncoated portion 102 u and coated portion 102 c which was hidden underneath the opaque character of insulation layer 106 .
  • insulation layer 106 is used to define the width of the first working electrode 100 , second working electrode 102 , and reference electrode 104 .
  • Insulation layer 106 further includes a cutout 108 which may have a T-shaped structure as shown in FIG. 4 to 6 .
  • Cutout 108 exposes a portion of first working electrode 100 , second working electrode 102 , and reference electrode 104 which can be wetted with liquid.
  • Cutout 108 further includes a distal cutout width W 11 , proximal cutout width W 12 , a distal cutout length L 14 and a proximal cutout length L 15 as illustrated in FIG. 5 and 6 .
  • Distal cutout width W 11 corresponds to the width of uncoated portion 102 u .
  • Distal cutout length L 14 is greater than the length uncoated portion 102 u .
  • Proximal cutout width W 12 and proximal cutout length L 15 forms a rectangular section which approximately exposes the width and length of first working electrode 100 , reference electrode 104 , and coated portion 102 c.
  • distal cutout width W 11 , proximal cutout width W 12 , distal cutout length L 14 and proximal cutout length L 15 may have a respective dimension of approximately 1.1, 0.7, 2.5, and 2.6 mm.
  • uncoated portion 102 u , reference electrode 104 , first working electrode 100 , and coated portion 102 c have a respective length of L 10 , L 12 , L 11 , and L 13 which may be about 0.7, 0.7, 0.4, and 0.4 mm.
  • Electrode spacing S 11 is a distance between uncoated portion 102 u and reference electrode 104 which may be between about 0.2 to 0.75 mm, and more preferably between 0.6 to 0.75 mm.
  • Electrode spacing S 10 is a distance between reference electrode 104 and first working electrode 100 ; and between coated portion 102 c and first working electrode 100 which may be about 0.2 mm.
  • electrode spacing S 11 is greater than S 10 to decrease the possibility of reagent dissolving and migrating to uncoated portion 102 u . Additionally, electrode spacing S 11 is greater than S 10 to decrease the possibility of reagent layer 110 being disposed on uncoated portion 102 u because of variations in the printing process.
  • the fourth through sixth layer which is successively disposed on strip 162 in the same manner as the first strip embodiment. The relative position and shape of the adhesive layer 66 , hydrophilic layer 68 , and top layer 40 are illustrated in FIG. 4 .
  • the C-shape of second working electrode 102 may be partially altered so that the order in which liquid would wet the electrodes would be uncoated portion 102 u , first working electrode 100 , reference electrode 104 , and then coated portion 102 c .
  • first working electrode 100 and coated portion 102 c would be equidistant from reference electrode 104 which is desirable from an IR drop perspective.
  • the electrodes are arranged so that the order in which liquid would wet the electrodes would be uncoated portion 102 u , reference electrode 104 , first working electrode 100 , and then coated portion 102 c .
  • coated portion 102 c is farther away from reference electrode 104 than the distance between first working electrode 100 and reference electrode 104 .
  • An algorithm may, therefore be used to calculate a corrected glucose current that is independent of interferences.
  • a constant potential is applied to the first and second working electrodes and a current is measured for both electrodes.
  • WE 2 G+I cov +I unc (Eq 2) where WE 2 is a current density at the second working electrode and I unc is a current density due to interferences at the portion of a working electrode not covered with reagent.
  • Uncoated portions 12 u can oxidize interferents, but not glucose because it is not coated with reagent layer 22 .
  • coated portion 12 c can oxidize glucose and interferents. Because it was experimentally found that uncoated portions 12 u oxidizes interferents in a manner proportional to the area of coated portion 12 c , it is possible to predict the proportion of interferent current measured overall at second working electrode 12 . This allows the overall current measured at second working electrode 12 to be corrected by subtracting the contribution of the interferent current.
  • the ratio of A unc :A cov may be between about 0.5:1 to 5:1, and is preferably about 3:1. More details describing this mathematical algorithm for current correction will be described in a later section.
  • the interferent oxidation current density measured at the coated portion may be different than the current density measured at the uncoated portion. This may be ascribed to a more efficient or less efficient oxidation of interferents at the coated portion.
  • the presence of a mediators may enhance the oxidation of interferences relative to the uncoated portion.
  • the presence of viscosity increasing substances such as hydroxyethyl cellulose may decrease the oxidation of interferences relative to the uncoated portion.
  • the interferent oxidation current density measured at the coated portion may be more or less than the uncoated portion.
  • Equation 1, 2, and 3a may be manipulated to derive an equation that outputs a corrected glucose current density independent of interferences.
  • the three equations (Equation 1, 2, and 3a) collectively have 3 unknowns which are G, I cov , and I unc .
  • Equation 1 can be rearranged to the following form.
  • G WE 1 ⁇ I cov (Eq 4)
  • I cov from Equation 3a can be substituted into Equation 4 to yield Equation 5.
  • G WE 1 - [ A cov A unc ⁇ I unc ] ( Eq ⁇ ⁇ 5 )
  • Equation 1 and Equation 2 can be combined to yield Equation 6.
  • Equation 7a outputs a corrected glucose current density G which removes the effects of interferences requiring only the current density output of the first and second working electrode, and a proportion of the coated to uncoated area of the second working electrode.
  • the proportion A cov A unc may be programmed into a glucose meter, in, for example, a read only memory.
  • the proportion A cov A unc may be transferred to the meter via a calibration code chip which would may account for manufacturing variations in A cov or A unc .
  • Equation 1 Equation 1 + Equation 7b
  • the corrected glucose current Equation 7a or 7b may be used by the meter only when a certain threshold is exceeded. For example, if WE 2 is about 10% or greater than WE 1 , then the meter would use Equation 7a or 7b to correct for the current output. However, if WE 2 is about 10% or less than WE 1 , the meter would simple take an average current value between WE 1 and WE 2 to improve the accuracy and precision of the measurement. The strategy of using Equation 7a or 7b only under certain situations where it is likely that a significant level of interferences are in the sample mitigates the risk of overcorrecting the measured glucose current. It should be noted that when WE 2 is sufficiently greater than WE 1 (e.g.
  • this is an indicator of having a sufficiently high concentration of interferents.
  • the first and second working electrodes are partially covered with the reagent layer in such a way that that the uncoated portions of the first and second working electrodes are different. This contrasts the previously described first and second test strip embodiments where the first working electrode is completely covered with the reagent layer.
  • FIG. 9 is a simplified plane view of a distal portion of a test strip 2000 according to yet another embodiment of the present invention wherein a reagent layer 22 is illustrated with the conductive layer and insulation layer 2002 such there are two working electrodes which have an uncoated portion.
  • Test strip 2002 is manufactured in a manner similar to test strip 62 except that there is a geometric change to cutout 18 as shown in FIG. 1 .
  • Test strip 2002 has the same substrate 50 , conductive layer 64 , reagent layer 22 , adhesive layer 66 , hydrophilic layer 68 , and top layer 40 as test strip 62 .
  • Test strip 2002 was modified to have a cutout 2004 which has a dumbbell like shape as illustrated in FIG. 9 .
  • the modified shape for cutout 2004 allows first working electrode 2008 to include a first coated portion 2008 c and an first uncoated portion 2008 u ; and second working electrode 2006 to include a second coated portion 2006 c and second uncoated portion 2006 u .
  • the two first uncoated portions 2008 u must have a different total area than the two second uncoated portions 2006 u.
  • FIG. 10 is a simplified plane view of a distal portion of a test strip 5000 according to still yet another embodiment of the present invention wherein a reagent layer 820 is illustrated with the conductive layer such there are two working electrodes which have an uncoated portion.
  • Test strip 5000 is manufactured in a manner similar to test strip 162 except that there is a geometric change to conductive layer 164 such that both a first working electrode 4002 and a second working electrode 4004 have a C-shape.
  • Test strip 5000 has the same substrate 50 , insulation layer 106 , reagent layer 110 , adhesive layer 66 , hydrophilic layer 68 , and top layer 40 as test strip 162 .
  • first working electrode 4002 to include a first coated portion 4002 c and an first uncoated portion 4002 u ; and second working electrode 4004 to include a second coated portion 4004 c and second uncoated portion 4004 u .
  • first uncoated portion 4002 u must have a different area than second uncoated portion 4004 u.
  • Test strips 2000 and 5000 have an advantage in that they may be easier to manufacture in regards to depositing the reagent layer with the required registration and also any subsequently deposited layers. Furthermore, both the first and second working electrodes will have to some extent the same chemical and electrochemical interactions with any interfering substances thus ensuring greater accuracy in the correction process. With both working electrodes having some level of uncoated area the same reactions will occur on both electrodes but to a different extent.
  • f 1 A cov1 A unc1
  • f 2 A cov1 A unc2
  • a unc1 is an uncoated area of the first working electrode
  • a unc2 is an uncoated area of the second working electrode
  • a cov1 is a coated area of the first working electrode
  • a cov2 is a coated area of the second working electrode.
  • One advantage of the present invention is the ability to use the first and second working electrode to determine that the sample receiving chamber has been sufficiently filled with liquid. It is an advantage of this invention in that the second working electrode not only corrects the interferent effect, but can also measure. glucose. This allows for a more accurate results because 2 glucose measurements can be averaged together while using only one test strip.
  • Test strips were prepared according to the first embodiment of the present invention as illustrated in FIG. 1 to 3 . These test strips were tested in blood having various concentrations of interferents. To test these strips, they were electrically connected to a potentiostat which has the means to apply a constant potential of 0.4 volts between the first working electrode and the reference electrode; and the second working electrode and the reference electrode. A sample of blood is applied to the sample inlet allowing the blood to wick into the sample receiving chamber and to wet first working electrode, second working electrode, and reference electrode. The reagent layer becomes hydrated with blood and then generates ferrocyanide which may be proportional to the amount of glucose and/or interferent concentration present in the sample. After about 5 seconds from the sample application to the test strip, an oxidation of ferrocyanide is measured as a current for both the first and second working electrode.
  • FIG. 11 shows the current responses of the first working electrode tested with 70 mg/dL glucose samples in blood spiked with varying levels of uric acid.
  • the uncorrected current at the first working electrode shows an increase in current that is proportional to the uric acid concentration.
  • the corrected current (depicted by triangles) which is processed by Equation 7a shows no effect from the increasing uric acid concentration.
  • FIG. 12 shows the current responses of the first working electrode tested with 240 mg/dL glucose samples in blood spiked with varying levels of uric acid.
  • the purpose of testing strips at 240 mg/dL glucose is to show that the correction algorithm of Equation 7a is also valid over a range of glucose concentrations.
  • the uncorrected current at the first working electrode shows an increase in current that is proportional to the uric acid concentration.
  • the corrected current shows no effect from the increasing uric acid concentration.
  • Equation 7a shows that the method of correcting the current output of the first working electrode using Equation 7a is effective in correcting for interferences.
  • Table 1 shows that the current correction in Equation 7a is effective for interferences with respect to acetaminophen, gentisic acid, and uric acid. Table 1 also shows the concentration range of the interferent which is normally found in blood. In addition, Table 1 also shows that the current correction in Equation 7a is effective at 240 mg/dL glucose concentration level.
  • FIG. 13 shows an exploded perspective view of a test strip 800 that is designed to lance a user's skin layer so as cause physiological fluid to be expressed and collected into test strip 800 in a seamless manner.
  • Test strip 800 includes a substrate 50 , a conductive layer 802 , an insulation layer 804 , a reagent layer 820 , an adhesive layer 830 , and a top layer 824 .
  • Test strip 800 further includes a distal end 58 and a proximal end 60 .
  • conductive layer 802 is the first layer disposed on substrate 50 .
  • Conductive layer 802 includes a second working electrode 806 , a first working electrode 808 , a reference electrode 810 , a second contact 812 , a first contact 814 , a reference contact 816 , a strip detection bar 17 , as shown in FIG. 13 .
  • the material used for conductive layer 802 and the process for printing conductive layer 802 is the same for both test strip 62 and test strip 800 .
  • Insulation layer 804 is the second layer disposed on substrate 50 .
  • Insulation layer 16 includes a cutout 18 which may have a rectangular shaped structure. Cutout 18 exposes a portion of second working electrode 806 , first working electrode 808 , and reference electrode 810 which can be wetted with a liquid.
  • the material used for insulation layer 804 and the process for printing insulation layer 804 is the same for both test strip 62 and test strip 800 .
  • Reagent layer 820 is the third layer disposed on substrate 50 , first working electrode 808 and reference electrode 810 .
  • the material used for reagent layer 820 and the process for printing reagent layer 820 is the same for both test strip 62 and test strip 800 .
  • Adhesive layer 830 is the fourth layer disposed on substrate 50 .
  • the material used for adhesive layer 830 and the process for printing adhesive layer 830 is the same for both test strip 62 and test strip 800 .
  • the purpose of adhesive layer 830 is to secure top layer 824 to test strip 800 .
  • top layer 824 may be in the form of an integrated lance as shown in FIG. 13 .
  • top layer 824 may include a lance 826 which is located at distal end 58 .
  • Lance 826 which may also be referred to as a penetration member, may be adapted to pierce a user's skin and draw blood into test strip 800 such that second working electrode 806 , first working electrode 808 , and reference electrode 810 are wetted.
  • Lance 826 includes a lancet base 832 that terminates at distal end 58 of the assembled test strip.
  • Lance 826 may be made with either an insulating material such as plastic, glass, and silicon, or a conducting material such as stainless steel and gold. Further descriptions of integrated medical devices that use an integrated lance can be found in International Application No. PCT/GB01/05634 and U.S. patent application Ser. No. 10/143,399.
  • lance 826 can be fabricated, for example, by a progressive die-stamping technique, as disclosed in the aforementioned International Application No. PCT/GBO1/05634 and U.S. patent application Ser. No. 10/143,399.
  • FIG. 14 is a simplified schematic showing a meter 900 interfacing with a test strip.
  • the following test strips may be suitable for use with meter 900 which are test strip 62 , test strip 162 , test strip 800 , test strip 2000 , test strip 3000 , or test strip 5000 .
  • Meter 900 has at least three electrical contacts that form an electrical connection to the second working electrode, the first working electrode, and the reference electrode.
  • second contact ( 13 , 103 , or 812 ) and reference contact ( 15 , 105 , or 816 ) connect to a first voltage source 910 ;
  • first contact ( 11 , 101 , or 814 ) and the reference contact ( 15 , 105 , or 816 ) connect to a second voltage source 920 .
  • first voltage source 910 applies a first potential E 1 between the second working electrode and the reference electrode; and second voltage source 920 applies a second potential E 2 between the first working electrode and the reference electrode.
  • first potential E 1 and second potential E 2 may be the same such as for example about +0.4 V.
  • first potential El and second potential E 2 may be different.
  • a sample of blood is applied such that the second working electrode, the first working electrode, and the reference electrode are covered with blood. This allows the second working electrode and the first working electrode to measure a current which is proportional to glucose and/or non-enzyme specific sources. After about 5 seconds from the sample application, meter 900 measures an oxidation current for both the second working electrode and the first working electrode.

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