CN100473983C - Method for reducing the effect of direct interference current in an electrochemical test strip - Google Patents
Method for reducing the effect of direct interference current in an electrochemical test strip Download PDFInfo
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- CN100473983C CN100473983C CNB2004800395335A CN200480039533A CN100473983C CN 100473983 C CN100473983 C CN 100473983C CN B2004800395335 A CNB2004800395335 A CN B2004800395335A CN 200480039533 A CN200480039533 A CN 200480039533A CN 100473983 C CN100473983 C CN 100473983C
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Abstract
This invention describes an electrochemical sensor which is adapted to reduce the effects of interfering compounds in bodily fluids when measuring an analyte in such fluids using an electrochemical strip (62). The sensor includes a substrate (50), a first and second working electrodes (10, 12), and a reference electrode (14). A reagent layer (22) is disposed on the electrodes such that, in one embodiment it completely covers all of the first working electrode, but only partially covers the second working electrode and, in a second embodiment, it only covers a portion of the first and the second working electrode. The portion of the working electrodes not covered by the reagent layer and is used to correct for the interference effect on the analyte measurement.
Description
Invention field
The present invention relates generally to reduce the method for interfering compound to the influence of the measurement undertaken by analyte measuring system, more particularly, relate to the method that reduces the influence of direct interference current in the glucose monitoring system that uses electrochemical test bar, described electrochemical test bar has the electrode that uncoated area is arranged.
Background of invention
Under many circumstances, electrochemical measurement system can have the oxidation current that increases owing to the oxidation of interfering compound common in the physiological fluid, and described interfering compound is for example paracetamol, ascorbic acid, cholerythrin, dopamine, gentianic acid, glutathione, levodopa, ethyldopa, tolazamide, orinase and uric acid.Therefore, by that part of oxidation current that reduces or elimination is produced by interfering compound, the accuracy that can improve glucose meters.Ideal situation is, the oxidation current that should not produce by any interfering compound, and whole like this oxidation current only depends on concentration of glucose.
Therefore, wish to improve at possible interfering compound for example in physiological fluid in the presence of common ascorbate, urate and the paracetamol accuracy of electrochemical sensor.For such electrochemical sensor, the example of analyte can comprise glucose, lactate and fructosamine.Though glucose is the main analyte of being discussed, it will be apparent to one skilled in the art that the present invention also can be used for other analyte.
Oxidation current can produce by several approach.Particularly, desired oxidation current is produced by the interaction of redox mediators with the analyte of being paid close attention to (for example glucose), and the oxidation current of not expecting is produced by the interfering compound oxidized with the interaction of redox mediators oxidized at electrode surface and that pass through usually.For example, some interfering compound (for example paracetamol) is oxidized at electrode surface.Other interfering compound (for example ascorbic acid) is by oxidized with the chemical reaction of redox mediators.In the glucose measurement system, the oxidation of interfering compound causes that concentration that the oxidation current of measurement had both depended on glucose also depends on the concentration of any interfering compound.Therefore, at interfering compound with the efficient oxidation same with glucose, and with respect to concentration of glucose, when the concentration of interfering compound was high, the measurement of concentration of glucose can improve to the contribution of total oxidation current by reducing or eliminating interfering compound.
A known strategy that reduces the influence of interfering compound is to use electronegative film to cover working electrode.As an example, can use for example NAFION of sulfonation fluorinated polymer
TMRepel all electronegative chemical substances.Generally speaking, most of interfering compound for example ascorbate and urate has negative charge, and therefore, electronegative film stops electronegative interfering compound to arrive electrode surface and oxidized on electrode surface.Yet for example paracetamol is not electronegative owing to some interfering compound, thereby and can pass through electronegative film, so this technology always the success.This technology can not reduce the oxidation current that the interaction owing to interfering compound and some redox mediators produces.For example the ferricyanide comes by electronegative film and electrode carries out electron exchange to use electronegative film also can stop some redox mediators commonly used on working electrode.
Another strategy that can be used for reducing the influence of interfering compound is to use the size Selection film at the working electrode top.As an example, can with 100 dalton's size exclusion films for example acetyl cellulose film cover working electrode, to get rid of molecular weight greater than 100 daltonian all chemical substances.The molecular weight of most of interfering compound is greater than 100 dalton, therefore is excluded and can not be oxidized on electrode.Yet such selection film makes that usually the manufacturing of test-strips is more complicated, and because the glucose of oxidation must increase Measuring Time by selecting film diffusion to arrive electrode.
Another strategy that can be used for reducing the influence of interfering compound is to use the redox mediators with suboxides reduction potential, for example oxidation-reduction potential for pact-300mV extremely+redox mediators of 100mV (when measuring) about saturated calomel electrode.Because redox mediators has the suboxides reduction potential, the voltage that imposes on working electrode also can be lower, and this has reduced interfering compound by the speed of working electrode oxidation.The example that has than the redox mediators of suboxides reduction potential comprises osmium bipyridyl complexes, ferrocene derivatives and quinone derivative.This tactful shortcoming is, the redox mediators that has than the suboxides reduction potential often is difficult to synthesize, and is unstable and have a low water solubility.
Another strategy that can be used for reducing the influence of interfering compound is to use the pseudo electrode that has been coated with redox mediators.In some cases, can also be with the oxidoreducing enzyme of pseudo electrode with inert protein or inactivation.The purpose of pseudo electrode is the redox mediators of the disturbed compound reduction of oxidation interfering compound and/or oxidation on electrode surface.In this strategy, will from total oxidation current of measuring at working electrode, deduct at the electric current of measuring on the pseudo electrode, to eliminate disturbing effect.This tactful shortcoming is that it needs test-strips to comprise can not be used to measure the other electrode of glucose and other electrical connection (being pseudo electrode).Comprise that pseudo electrode is to use electrode in the glucose measurement system inefficiently.
Summary of the invention
The present invention relates to when using electrochemical sensor to come the check and analysis thing, reduce the method for chaff interference influence.The electrochemical sensor that can be used for the inventive method comprises substrate, first and second working electrode and contrast electrode at least.Reagent layer is arranged on the electrode, makes reagent layer cover whole districts of first working electrode fully, and only part covers second working electrode.In the methods of the invention, use the oxidation current that on the part that is not covered of second working electrode, produces to proofread and correct the influence of chaff interference to glucose measurement by reagent layer.
The present invention also comprises the method that reduces the interference in the electrochemical sensor, comprises the following steps: to measure first oxidation current on first working electrode, and wherein first working electrode is covered by reagent layer; Second oxidation current of measurement on second working electrode, wherein reagent layer only partly covers second working electrode; The correction oxidation current of the concentration of preselected analyte (for example glucose) with calculating representative.In this calculates, use the area coverage of second working electrode and the ratio of area coverage not to eliminate the influence of chaff interference to oxidation current.More particularly, can use following formula to come the calculation correction current value,
Wherein G is a correcting current density, WE
1Be the not correcting current density on first working electrode, WE
2Be the not correcting current density on second working electrode, A
CovBe the spreading area of second working electrode, A
UncIt is the uncoated area of second working electrode.
In can be used for an embodiment of electrochemical test bar of the present invention, electrochemical glucose test strips comprises first and second working electrode, wherein first working electrode is covered fully by reagent layer, and second working electrode only covered by the reagent layer segment.Therefore, second working electrode has reagent spreading area and uncoated area.Reagent layer for example can comprise for example glucose oxidase and the redox mediators ferricyanide for example of oxidoreducing enzyme.First working electrode will have the stack of two oxidation current sources, and one from glucose, and another is from chaff interference.Similarly, second working electrode will have the stack of three current sources, these three current sources respectively from glucose, at chaff interference on the reagent coated portion and the chaff interference on uncoated part.Owing in this zone, do not have reagent, thus the uncoated part of second working electrode oxidation chaff interference only, and oxidizing glucose not.The oxidation current of measuring on the uncoated part of second working electrode can be owing to estimating total interferent oxidation current and calculating the correction oxidation current of having eliminated the chaff interference influence.
In another test-strips embodiment in can be used for the inventive method, electrochemical glucose test strips comprises first and second working electrode, and wherein first is only covered by the reagent layer segment with second working electrode.Therefore, in this embodiment, first and second working electrode all have reagent coated portion and uncoated part.First of first working electrode not area coverage and second working electrode second not area coverage be different.The oxidation current that use is measured on the uncoated part of first and second working electrode is estimated the interferent oxidation current and the calculation correction glucose current of uncoated part.
The present invention also comprises the method that reduces the interference in the electrochemical sensor, comprises the following steps: to measure first oxidation current on first working electrode, and wherein first working electrode is covered by the reagent layer segment; Second oxidation current of measurement on second working electrode, wherein reagent layer only partly covers second working electrode; The correction oxidation current of the concentration of preselected analyte (for example glucose) with calculating representative.In this calculates, use the area coverage of first and second working electrode and the ratio of area coverage not to eliminate the influence of chaff interference to oxidation current.More particularly, can use following formula to come the calculation correction current value,
F wherein
1Equal A
Cov1/ A
Unc1f
2Equal A
Cov2/ A
Unc2A
Unc1It is the uncoated area of first working electrode; A
Unc2It is the uncoated area of second working electrode; A
Cov1It is the spreading area of described first working electrode; A
Cov2It is the spreading area of second working electrode; G is the correcting current value; WE
1Be the not correcting current density on first working electrode, and WE
2Be the not correcting current density on second working electrode.
The accompanying drawing summary
By the following detailed description that provides exemplary, can understand the features and advantages of the present invention better, wherein used principle of the present invention and accompanying drawing:
Fig. 1 is the exploded perspective illustration of test-strips according to an embodiment of the invention.
Fig. 2 is the simplification plan view of the distal portions of test-strips, and described test-strips is the test-strips according to embodiment of the present invention shown in Figure 1, and comprises conductive layer and insulation course.
Fig. 3 is the simplification plan view according to the distal portions of the test-strips of embodiment of the present invention shown in Figure 1, has wherein shown the position of reagent layer and insulation course and conductive layer.
Fig. 4 is the exploded perspective illustration of test-strips according to another embodiment of the invention.
Fig. 5 is the simplification plan view of the distal portions of test-strips, and described test-strips is the test-strips according to embodiment of the present invention shown in Figure 4, and comprises conductive layer and insulation course.
Fig. 6 is the simplification plan view according to the distal portions of the test-strips of embodiment of the present invention shown in Figure 4, has wherein shown reagent layer and insulation course and conductive layer.
Fig. 7 is the simplification plan view according to the distal portions of the test-strips of embodiment of the present invention shown in Figure 4, has wherein shown reagent layer and conductive layer.
Fig. 8 is the simplification plan view of the distal portions of test-strips according to another embodiment of the invention, has wherein shown reagent layer, and it has the conductive layer that helps to reduce IR potential drop effect.
Fig. 9 is the simplification plan view of the distal portions of test-strips according to another embodiment of the invention, has wherein shown reagent layer and conductive layer and insulation course, has two working electrodes with uncoated part like this.
Figure 10 is the simplification plan view of the distal portions of test-strips according to another embodiment of the invention, has wherein shown reagent layer and conductive layer and insulation course, has two working electrodes with uncoated part like this.
Figure 11 has represented the electric current on first working electrode of the test-strips of design according to the present invention, and measurement is to use the blood sample of the 70mg/dL glucose that has added the varying level uric acid to carry out.
Figure 12 has represented the electric current on first working electrode of the test-strips of design according to the present invention, and measurement is to use the blood sample of the 240mg/dL glucose that has added the varying level uric acid to carry out.
Figure 13 is the exploded perspective illustration with test-strips of integral knife blade.
Figure 14 is the simplified schematic diagram illustrating that shows the measuring instrument that is connected with test-strips, and described test-strips has first contact point (contact) of being arranged on the substrate, second contact point and reference contact point.
Detailed Description Of The Invention
The present invention includes the optionally test-strips and the method that are used to improve the electrochemical glucose measuring system.
Fig. 1 is the exploded perspective illustration of the test-strips of first embodiment according to the present invention.In embodiment of the present invention shown in Figure 1, be used for measuring body fluid for example the electrochemical test bar 62 of blood or interstitial fluid concentration of glucose comprise first working electrode 10 and second working electrode 12, wherein first working electrode 10 is covered fully by reagent layer 22, and second working electrode 12 only covered by reagent layer 22 parts.Therefore, second working electrode has reagent coated portion and uncoated part.Reagent layer 22 for example can comprise for example ferricyanide of oxidoreducing enzyme such as glucose oxidase and redox mediators.Because the ferricyanide is the oxidation-reduction potential that has about 400mV on the carbon electrode (when measuring with respect to saturated calomel electrode), so introducing body fluid for example blood can produce significant chaff interference oxidation by redox mediators and/or working electrode, thereby produce the oxidation current of not expecting significantly.Therefore, the oxidation current of measuring on first working electrode 10 will be the stack of oxidation current sources: because the oxidation current of first expectation that the oxidation of glucose produces and second oxidation current of not expecting by the chaff interference generation.The oxidation current of measuring on second working electrode 12 also will be the stack of oxidation current sources: the oxidation current of first expectation that produces owing to the oxidation of glucose, at second oxidation current of not expecting that produces by chaff interference on the cover part of second working electrode 12 and the 3rd oxidation current that on the unmasked portion of second working electrode 12, produces by chaff interference.The uncoated part of second working electrode 12 is the oxidation chaff interference only, and oxidizing glucose not, because on the uncoated part of second working electrode 12, do not have reagent.Because oxidation current of measuring on the uncoated part of second working electrode 12 and glucose are irrelevant, and the uncoated area of two working electrodes 12 is known, so can calculate the interferent oxidation current of the uncoated part of second working electrode 12.Thereby, the interferent oxidation current of the uncoated part of second working electrode 12 that use to calculate and know the area of first working electrode 10 and the area of the coated portion of second working electrode 12 can calculate the corrected glucose current of having eliminated the influence of the interfering compound of oxidation on electrode.
Fig. 1 is the exploded perspective illustration of the test-strips 62 of first embodiment according to the present invention.Test-strips 62 as shown in Figure 1 can be printed step continuously by a series of 6 and produce, and these steps are that 6 layer materials are placed on the substrate 50.Can print by for example screen cloth these 6 are deposited upon on the substrate 50.In one embodiment of the invention, these 6 layers can comprise conductive layer 64, insulation course 16, reagent layer 22, bonding coat 66, hydrophilic layer 68 and top layer 40.Conductive layer 64 can further comprise first working electrode 10, second working electrode 12, contrast electrode 14, first contact point 11, second contact point 13, reference contact point 15 and test-strips check-out console 17.Insulation course 16 can further comprise otch (cutout) 18.Bonding coat 66 can further comprise first adhesive cushion 24, second adhesive cushion 26 and the 3rd adhesive cushion 28.Hydrophilic layer 68 can further comprise first hydrophilic film 32 and second hydrophilic film 34.Top layer 40 can further comprise transparent part 36 and opaque section 38.As shown in Figure 1, test-strips 62 has first side 54 and second side 56, electrode distally 58 and electrode nearside 60.Following chapters and sections will be described each layer of test-strips 62 in more detail.
In one embodiment of the invention, substrate 50 is for example plastics, glass, potteries etc. of electrically insulating material.In a preferred embodiment of the invention, substrate 50 can be for example nylon, polycarbonate, polyimide, Polyvinylchloride, tygon, polypropylene, PETG or polyester of plastics.More particularly, polyester can be for example to be produced by DuPont Teijin Films
ST328.Substrate 50 can also comprise acrylic acid coatings, coating be coated in improve on one or two side the China ink bonding.
The ground floor that is deposited on the substrate 50 is a conductive layer 64, and this layer comprises first working electrode 10, second working electrode 12, contrast electrode 14 and test-strips check-out console 17.According to the present invention, can come for example conductive carbon ink of deposition materials with screen cloth with qualification geometry shown in Figure 1 with latex pattern.Contrast electrode 14 can also be counter electrode, contrast electrode/counter electrode or accurate contrast electrode.Can print, take turns the printing of commentaries on classics notch board, sputter, evaporation, electroless spraying, ink-jet, distillation, chemical vapor deposition etc. by screen cloth is deposited on conductive layer 64 on the substrate 50.The suitable material that can be used for conductive layer 64 is the tin oxide of Au, Pd, Ir, Pt, Rh, stainless steel, doping, carbon etc.In one embodiment of the invention, carbon ink layer can have the 1-100 micron, more especially 5-25 micron, even more especially about 13 microns height.The height of conductive layer can be according to the resistance of needed conductive layer and the conductivity of electrolyte materials that is used to print conductive layer change.
Second layer that is deposited on the substrate 50 is insulation course 16.As shown in Figure 1, insulation course 16 is deposited at least a portion conductive layer 64.Fig. 2 is the simplification plan view of the distal portions of test-strips 62, this figure first working electrode 10, second working electrode 12 and contrast electrode 14 position with respect to insulation course 16 of having given prominence to the key points.Insulation course 16 also comprises otch (cutout) 18, and it can have T-shape structure as illustrated in fig. 1 and 2.Otch 18 has exposed can be by first working electrode of the wetting part of liquid 10, second working electrode 12 and contrast electrode 14.Otch 18 further comprises distal incision width W 1, near-end kerf width W2, distal incision length L 4 and near-end incision length L5.As shown in Figure 2, distal incision width W 1 is corresponding with the width of first working electrode 10 and contrast electrode 14.Distal incision length L 4 pairing length are greater than the length sum of first working electrode 10 with contrast electrode 14.Near-end kerf width W2 and near-end incision length L5 form and have exposed the width of second working electrode 12 and the rectangle tangent plane of length.According to the present invention, distal incision width W 1, near-end kerf width W2, distal incision length L 4 and near-end incision length L5 can have about 0.7,1.9,3.2 and the size of 0.43mm respectively.In one embodiment of the invention, first working electrode 10, contrast electrode 14 and second working electrode 12 have length L 1, L2 and L3 respectively, and they can be respectively about 0.8,1.6 and 0.4mm.Electrode separation S1 is the distance between first working electrode 10 and the contrast electrode 14; And the distance between contrast electrode 14 and second working electrode 12, it can be about 0.4mm.
The 3rd layer that is deposited on the substrate 50 is reagent layer 22.As shown in Figure 1, reagent layer 22 is arranged at least a portion of conductive layer 64 and insulation course 16.Fig. 3 is the simplification plan view of distal portions of the test-strips 62 of first embodiment according to the present invention, this figure position of reagent layer 22 with respect to first working electrode 10, second working electrode 12, contrast electrode 14 and insulation course 16 of having given prominence to the key points.Reagent layer 22 can be a rectangle as shown in figs. 1 and 3, and this rectangle has reagent width W 3 and reagent length L 6.In one embodiment of the invention, reagent width W 3 can be about 1.3mm, and reagent length L 6 can be about 4.7mm.In another embodiment of the invention, reagent layer 22 has enough big width W 3 and length L 6, makes reagent layer 22 cover first working electrode 10 and contrast electrode 14 fully.Yet reagent layer 22 width W 3 of a size suitable and length L 6 make second working electrode do not covered fully by reagent layer 22.In such scheme, as shown in Figure 3, second working electrode 12 has coated portion 12c and uncoated part 12u.Uncoated part 12u can become the shape of two rectangles, and wherein uncoated part 12u has wing width W 4 and corresponding to the length of second working electrode length L 3.As limiting examples, wing width W 4 can be about 0.3mm.In one embodiment of the invention, reagent layer 22 can comprise for example glucose oxidase or PQQ-glucose dehydrogenase (wherein PQQ is the acronym of pyrrolo--quinoline-quinone) and the redox mediators ferricyanide for example of oxidoreducing enzyme.
The 4th layer that is deposited on the substrate 50 is bonding coat 66, and it comprises first adhesive cushion 24, second adhesive cushion 26 and the 3rd adhesive cushion 28.The wall of first adhesive cushion 24 and second adhesive cushion 26 formation sample receiving chamber.In one embodiment of the invention, first adhesive cushion 24 and second adhesive cushion 26 can be arranged on the substrate 50, make these two adhesive cushions not contact reagent layer 22.Do not need therein to reduce in another embodiment of the invention of test-strips volume, first adhesive cushion 24 and/or second adhesive cushion 26 can be arranged on the substrate 50, make itself and reagent layer 22 overlapping.In one embodiment of the invention, bonding coat 66 has the height of about 70-110 micron.Bonding coat 66 can comprise bilateral contact adhesive, UV cure adhesive, heat-activated adhesive, hot setting adhesive or other bonding agent well known by persons skilled in the art.As limiting examples, bonding coat 66 can be printed contact adhesive by screen cloth and form, and described bonding agent is a water based acrylic copolymer contact adhesive for example, and it is available from Iape Specialties LTD in Tring, Herts, United Kingdom (part#A6435).
The 5th layer that is deposited on the substrate 50 is hydrophilic layer 68, and as shown in Figure 1, it comprises first hydrophilic film 32 and second hydrophilic film 34.Hydrophilic layer 68 forms " top, chamber " of sample receiving chamber." sidewall " of sample receiving chamber and " base plate " form by a part of bonding coat 66 and substrate 50 respectively.As limiting examples, hydrophilic layer 68 can be the optional transparent polyester of possess hydrophilic property antifogging coating, for example available from those of 3M.In the design of test-strips 62, use the water wettability character of coating, because it helps liquid filling in the sample receiving chamber.
The 6th layer that is deposited on the substrate 50 is top layer 40, and as shown in Figure 1, it comprises transparent part 36 and opaque section 38.According to the present invention, top layer 40 is included in the polyester of a side with the contact adhesive coating.Top layer 40 has opaque section 38, and when blood was below transparent part 36, it helped the user to observe high-contrast.This makes the user enough to be full of with eyesight validation sample receiving chamber.After with test-strips 62 complete laminations, it is cut along cut line A-A ', in this process, produced sample inlet 52 as shown in Figure 3.
First test-strips embodiment shown in Fig. 1-3 can have the shortcoming of possibility, because reagent layer 22 can dissolve in liquid sample, and the reagent layer that a part is dissolved moves on the uncoated part 12u of second working electrode 12.If such situation takes place, uncoated part 12u will also measure the oxidation current that also is directly proportional with concentration of glucose.This will reduce the ability of using mathematical algorithm to eliminate the chaff interference oxidation affects.In another embodiment of the invention, reagent layer 22 should be designed to dissolve in the mode of not moving on the uncoated part 12u.For example, can be with reagent layer 22 and first working electrode 10, second working electrode 12 and contrast electrode 14 chemical bond, perhaps can have the thickening agent that the migration of the reagent layer 22 of dissolving can be reduced to minimum degree.
Another embodiment of the invention as shown in Figure 4, embodiment shown in Figure 4 has reduced the migration of reagent on the uncoated part of second working electrode of dissolving, and in some cases, minimum degree is reduced in this migration.In this embodiment, as shown in Figure 4, second working electrode 102 has C-shape geometric configuration, and wherein 2 of second working electrode 102 discontinuous parts are exposed by otch 108.According to the present invention, as shown in Figure 6, reagent layer 110 only is arranged on second working electrode 102 of a part, to eliminate uncoated part 102u and coated portion 102c.It is 52 adjacent that uncoated part 102u and sample enter the mouth.Coated portion 102c is adjacent with first working electrode 100.When in the sample of the test-strips 162 that liquid the is applied to assembling inlet 52, liquid will enter the mouth from sample and 52 flow on the coated portion 102c, all be covered by liquid until all electrodes.By with the location arrangements of uncoated part 102u in the upstream of flow of liquid, this has almost completely prevented reagent layer 110 dissolvings and has moved on the uncoated part 102u.This makes mathematical algorithm can accurately eliminate the influence of chaff interference to the oxidation current of measurement.
Fig. 4 is the exploded perspective illustration of test-strips 162.Test-strips 162 is to make according to the mode that is similar to test-strips 62, but conductive layer 164, insulation course 106 and reagent layer 110 has been made the change of geometry or aspect, position.For second embodiment of the present invention, substrate 50, bonding coat 66, hydrophilic layer 68 and top layer 40 are identical with first test-strips embodiment.Test-strips 162 has first side 54 and second side 56, electrode distally 58 and electrode nearside 60.Shall also be noted that of the present invention first can comprise the parts with similar structures with second test-strips embodiment, these parts are to represent with identical numbering and title.If the like of each test-strips embodiment is structurally different, these parts can have same names, but provide with different unit numbers.Following chapters and sections will be described each layer of test-strips 162 in more detail.
For test-strips embodiment shown in Figure 4, first layer that is arranged on the substrate 50 is a conductive layer 164, and it comprises first working electrode 100, second working electrode 102, contrast electrode 104, first contact point 101, second contact point 103 and reference contact point 105 and test-strips check-out console 17.According to the present invention, can come for example conductive carbon ink of deposition materials with screen cloth with qualification geometry shown in Figure 4 with latex pattern.First contact point 101, second contact point 103 and reference contact point 105 can be used for being electrically connected with measuring instrument.This makes measuring instrument logical by first contact point 101, second contact point 103 and reference contact point 15 and first working electrode 100, second working electrode 102 and contrast electrode 104 Electricity Federations respectively.
In Fig. 4, second layer that is deposited on the substrate 50 is insulation course 106.As shown in Figure 4, insulation course 106 is deposited at least a portion conductive layer 164.Fig. 5 is the simplification plan view of the distal portions of test-strips 162, this figure first working electrode 100, second working electrode 102 and contrast electrode 104 position with respect to insulation course 106 of having given prominence to the key points.
In Fig. 4, the 3rd layer that is deposited on the substrate 50 is reagent layer 110, and as shown in Figure 6, reagent layer 110 is arranged at least a portion of conductive layer 164 and insulation course 106.Fig. 6 is the simplification plan view of distal portions of the test-strips 162 of second embodiment according to the present invention, this figure position of reagent layer 110 with respect to first working electrode 100, second working electrode 102, contrast electrode 104 and insulation course 106 of having given prominence to the key points.Reagent layer 110 can be a rectangle, and this rectangle has reagent width W 13 and reagent length L 16.In one embodiment of the invention, reagent width W 13 can be about 1.3mm, and reagent length L 16 can be about 3.2mm.In a preferred embodiment of the invention, reagent layer 110 has enough big width W 13 and length L 16, makes reagent layer 110 cover first working electrode 100, coated portion 102c and contrast electrode 104 fully, but does not cover uncoated part 102u.
Fig. 7 is the simplification plan view according to the distal portions of the test-strips of embodiment of the present invention shown in Figure 4, has wherein shown reagent layer and conductive layer.Different with Fig. 6, Fig. 7 does not demonstrate insulation course 106.This help has confirmed the conductive relation between uncoated part 102u and the coated portion 102c, its be hidden in insulation course 106 opaque symbol below.
For test-strips embodiment shown in Figure 4, insulation course 106 is used to limit the width of first working electrode 100, second working electrode 102, contrast electrode 104.Insulation course 106 also comprises otch 108, and it can have the T-shape structure shown in Fig. 4-6.Otch 108 has exposed can be by first working electrode of the wetting part of liquid 100, second working electrode 102 and contrast electrode 104.As illustrated in Figures 5 and 6, otch 108 further comprises distal incision width W 11, near-end kerf width W12, distal incision length L 14 and near-end incision length L15.Distal incision width W 11 is corresponding with the width of uncoated part 102u.Distal incision length L 14 is greater than the length of uncoated part 102u.Near-end kerf width W12 and near-end incision length L15 form and have exposed the width of first working electrode 100, contrast electrode 104 and coated portion 102c and the rectangle tangent plane of length substantially.
According to the present invention, distal incision width W 11, near-end kerf width W12, distal incision length L 14 and near-end incision length L15 can have about 1.1,0.7,2.5 and 2 respectively.The size of 6mm.
In the embodiment of Fig. 4, uncoated part 102u, contrast electrode 104, first working electrode 100 and coated portion 102c have length L 10, L12, L11 and L13 respectively, and they can be respectively about 0.7,0.7,0.4 and 0.4mm.Electrode separation S11 is the distance between uncoated part 102u and the contrast electrode 104, and it can be the more preferably about 0.6-0.7mm of about 0.2-0.75mm.Electrode separation S10 is the distance between contrast electrode 104 and first working electrode 100; And the distance between coated portion 102c and first working electrode 100, it can be about 0.2mm.Should be noted that electrode separation S11 greater than electrode separation S10, to reduce agent dissolves and the possibility of moving on the uncoated part 102u.In addition, electrode separation S11 is greater than electrode separation S10, makes reagent layer 110 be arranged in possibility on the uncoated part 102u to reduce owing to print difference in the processing.The the 4th to the 6th layer is deposited on the test-strips 162 successively in the mode identical with first test-strips embodiment.The relative position of bonding coat 66, hydrophilic layer 68 and top layer 40 and shape are as shown in Figure 4.
In embodiment of the present invention shown in Figure 8, can partly change the C-shape of second working electrode 102, make that the order of liquid wetting electrode is uncoated part 102u, first working electrode 100, contrast electrode 104 and coated portion 102c.In another form, first working electrode 100 and coated portion 102c equate that with the distance of contrast electrode 104 this expects from IR potential drop angle.In second test-strips embodiment shown in Figure 7 (being test-strips 162), the arrangement of electrode makes that the order of liquid wetting electrode is uncoated part 102u, contrast electrode 104, first working electrode 100 and coated portion 102c.For test-strips 162, the distance between coated portion 102c and the contrast electrode 104 is greater than the distance between first working electrode 100 and the contrast electrode 104.
Therefore, can use an algorithm to calculate the corrected glucose current of interference-free thing influence.After applying sample to test-strips, apply constant current potential for first working electrode and second working electrode, and measure the electric current of these two electrodes.Reagent covers on first working electrode of entire electrode area therein, can use following formula to describe the composition that oxidation current is made contributions,
WE
1=G+I
Cov(formula 1)
WE wherein
1Be the current density on first working electrode, G is the current density that produces owing to glucose of interference-free thing influence, and I
CovIt is the current density that produces owing to the chaff interference on the working electrode part that is covered by reagent.
On second working electrode that is partly covered by reagent, can use following formula to describe the composition that oxidation current is made contributions,
WE
2=G+I
Cov+ I
Unc(formula 2)
WE wherein
2Be the current density on second working electrode, I
UncIt is the current density that produces owing to the chaff interference on the working electrode part that is not covered by reagent.Can first with second working electrode on use different reagent spreading areas to obtain other embodiment of the present invention, but formula must be considered different uncoated areas.
In order to reduce the influence of chaff interference, the interference current on the coated portion that is described in second working electrode and the formula of the relation between the interference current on the uncoated part of second working electrode have been worked out.Estimation is approximately identical with the current density of measuring on uncoated part in the interferent oxidation current density of measuring on the coated portion.Further describe this relation by following formula,
A wherein
CovThe area of second working electrode that is covered by reagent, and A
UncIt is the area of second working electrode not covered by reagent.
Should be noted that uncoated part 12u and coated portion 12c can have is appointed as A separately
UncAnd A
CovArea.The oxidable chaff interference of uncoated part 12u, but oxidizing glucose not be not because be coated with reagent layer 22 above it.On the contrary, oxidable glucose of coated portion 12c and chaff interference.Because found through experiments, the mode oxidation chaff interference that uncoated part 12u is directly proportional with the area with coated portion 12c is so can the predicted interference electric current account for the ratio of the total current of measuring on second working electrode 12.This makes the total current of measuring on second working electrode 12 to be corrected by deducting interference current.In one embodiment of the invention, A
Unc: A
CovRatio can be about 0.5: 1-5: 1, and be preferably about 3: 1.More detailed description about this mathematical algorithm of current correction will be described in the chapters and sections below.
In another embodiment of the invention, can be different in the interferent oxidation current density of measuring on the coated portion with the current density of on uncoated part, measuring.This may be because, the chaff interference oxidation that efficient is higher or efficient is lower on coated portion.In one case, with respect to uncoated part, the redox mediators of existence can improve the oxidation of chaff interference.In another case, with respect to uncoated part, the material of the increase viscosity of existence for example hydroxyethyl cellulose can reduce the chaff interference oxidation.According to composition included in the reagent layer that second working electrode partly is coated with, the interferent oxidation current of measuring on coated portion can be bigger or little than uncoated part.This character can be undertaken phenomenological modelization by formula 3a is rewritten into following form,
I
Cov=f * I
Unc(formula 3b)
Wherein f is a correction factor, and it has introduced the influence of the chaff interference oxidation efficiency of coated portion to uncoated part.
In one embodiment of the invention, can operational formula 1,2 and the derive formula of the corrected glucose current density that can export interference-free thing influence of 3a.Should be noted that these three formula ( formula 1,2 and 3a) have 3 unknown numbers altogether, these 3 unknown numbers are G, I
CovAnd I
UncYet, formula 1 can be rearranged into following form.
G=WE
1-I
Cov(formula 4)
Next, can the I of formula 3a will be derived from
CovBe substituted in the formula 4, obtain formula 5.
Next, formula 1 and formula 2 can be merged, obtain formula 6.
I
Unc=WE
2-WE
1(formula 6)
Next, can the I of formula 6 will be derived from
UncBe substituted in the formula 5, obtain formula 7a.
Formula 7a has exported corrected glucose current density G, glucose current density G has eliminated the influence of chaff interference, this formula only needs the current density output from first working electrode and second working electrode, and the ratio of the spreading area of second working electrode and uncoated area.In one embodiment of the invention, can be with ratio A
Cov/ A
UncBe programmed in the glucose meters, for example be programmed in the ROM (read-only memory) of measuring instrument.In another embodiment of the invention, can be with ratio A
Cov/ A
UncPass to measuring instrument by the calibration code chip, described calibration code chip can be eliminated A
CovOr A
UncIn manufacturing variation.
In another embodiment of the invention, when the interferent oxidation current density of the interferent oxidation current density of coated portion and uncoated part not simultaneously, can use formula 1,2 and 3b.Under these circumstances, derive another updating formula 7b as follows.
G=WE
1-{ f * (WE
2-WE
1) (formula 7b)
In another embodiment of the invention, have only when surpassing certain threshold value, measuring instrument just can use corrected glucose current formula 7a or 7b.For example, if WE
2Compare WE
1Go out greatly more than about 10% or 10%, then measuring instrument will use formula 7a or 7b to proofread and correct output current.Yet, if WE
2Compare WE
1Go out greatly below about 10% or 10%, then measuring instrument will be got WE simply
1With WE
2Between average current value, improve the accuracy and the degree of accuracy of measurement.The danger of only under wherein there is the situation of interfering compound of the level of signifiance in the sample in some, using the strategy of formula 7a or 7b to alleviate the glucose current excessive correction of measuring.Should be noted that and work as WE
2Compare WE
1When (for example going out about 20% or more greatly), this is to have the very indication of the interfering compound of high concentration when enough big.Under these circumstances, may wish output error message rather than dextrose equivalent, because very high-caliber chaff interference can cause breaking of formula 7a or 7b accuracy.
In the embodiment of the present invention shown in Fig. 9 and 10, first and second working electrode are covered by the reagent layer segment by this way, make that the uncoated part of first and second working electrode is different.This with first working electrode wherein by reagent layer cover fully above-mentioned first are different with second test-strips embodiment.
Fig. 9 is the simplification plan view of the distal portions of test-strips 2000 according to another embodiment of the invention, has wherein shown reagent layer 22 and conductive layer and insulation course 2002, and two working electrodes with uncoated part are arranged.Test-strips 2000 is to prepare according to the mode that is similar to test-strips 62, but otch shown in Figure 1 18 has been made the geometry change.Test-strips 2002 has identical substrate 50, conductive layer 64, reagent layer 22, bonding coat 66, hydrophilic layer 68 and top layer 40 with test-strips 62.Change test-strips 2002, make it have otch 2004, this otch has dumbbell sample shape as shown in Figure 9.Shape to the change of otch 2004 makes first working electrode 2008 comprise first coated portion 2008c and first uncoated part 2008u; Second working electrode 2006 comprises second coated portion 2006c and second uncoated part 2006u.In order to allow test-strips 2000 reduce the influence of chaff interference effectively, first uncoated part 2008u must have and second total area that uncoated part 2006u is different.
Figure 10 is the simplification plan view of the distal portions of test-strips 5000 according to another embodiment of the invention, has wherein shown reagent layer 820 and conductive layer, and two working electrodes with uncoated part are arranged.Test-strips 5000 is to prepare according to the mode that is similar to test-strips 162, but conductive layer 164 has been made the geometry change, makes first working electrode 4002 and second working electrode 4004 all have the C-shape.Test-strips 5000 has same substrate 50, insulation course 106, reagent layer 110, bonding coat 66, hydrophilic layer 68 and top layer 40 with test-strips 162.The geometry of this change makes first working electrode 4002 comprise first coated portion 4002c and first uncoated part 4002u; Second working electrode 4004 comprises second coated portion 4004c and second uncoated part 4004u.In order to allow test-strips 2000 reduce the influence of chaff interference effectively, first uncoated part 4002u must have and second area that uncoated part 4004u is different.
Test-strips 2000 and 5000 advantage are that aspect the layer that reagent layer is deposited to desired position and any deposition subsequently, they can be easy to produce.In addition, first will have identical and chemistry any chaff interference with second working electrode on some degree and galvanochemistry interacts, and has guaranteed to have bigger accuracy thus in bearing calibration.Because two uncoated areas that working electrode all has some level, identical still in various degree reaction will take place on two electrodes.Formula 7a is made simple change, and following formula 7c can be used as the updating formula of glucose,
F wherein
1=A
Cov1/ A
Unc1, f
2=A
Cov2/ A
Unc2, A
Unc1The uncoated area of=first working electrode, A
Unc2The uncoated area of=the second working electrode, A
Cov1The spreading area of=first working electrode, and A
Cov2The spreading area of=the second working electrode.
An advantage of the invention is, can use first and second working electrode to determine that the sample receiving chamber enough is full of by liquid.An advantage of the invention is that second working electrode not only proofreaied and correct the chaff interference influence, but also can measure glucose.This can obtain can accurately the result, because under the situation of using a test-strips, 2 glucose measurements can be averaged.
Prepare test-strips according to first embodiment of the present invention shown in Fig. 1-3.In blood, measure these test-strips with variable concentrations chaff interference.In order to measure these test-strips, they are electrically connected with potentiostat, potentiostat has parts between first working electrode and contrast electrode 810; And apply 0.4 volt constant potential between second working electrode and the contrast electrode.Apply blood sample to sample inlet, allow blood soak and be drawn onto in the sample receiving chamber, and wetting first working electrode, second working electrode and contrast electrode.Reagent layer becomes by the blood hydration, produces ferrocyanide then, and ferrocyanide can be directly proportional with the amount and/or the interferent concentration of the glucose that exists in the sample.Applying sample to test-strips after 5 seconds, measurement is as the oxidation of the ferrocyanide of the electric current of first working electrode and second working electrode.
Figure 11 has represented the current response on first working electrode, and measurement is to use the blood sample of the 70mg/dL glucose that has added the varying level uric acid to carry out.Not correcting current (representing with square) on first working electrode shows the electric current that is directly proportional with uric acid concentration to be increased.Yet, the influence that the correcting current of handling by formula 7a (representing with triangle) shows the uric acid concentration that is not increased.
Figure 12 has represented the current response on first working electrode, and measurement is to use the blood sample of the 240mg/dL glucose that has added the varying level uric acid to carry out.Purpose in 240mg/dL glucose assays test-strips is to show, the correcting algorithm of formula 7a also is effective under the concentration of glucose of certain limit.Similar with Figure 11, the not correcting current (representing with square) on first working electrode shows the electric current that is directly proportional with uric acid concentration to be increased.Yet, the influence that correcting current (representing with triangle) shows the uric acid concentration that is not increased.
In order to show that the method for proofreading and correct the chaff interference electric current is applicable to multiple chaff interference, except uric acid, also the paracetamol of usefulness variable concentrations level and gentianic acid are measured the test-strips according to the embodiment structure of Fig. 1.In order quantitatively to determine the size of this influence, will be greater than 10% (for glucose level〉70mg/dL) or 7mg/dL (change for the glucose output of glucose level≤70mg/dL) and to be defined as remarkable interference.Table 1 shows, compares with the test-strips of correcting current reaction assay by using formula 7a, and the not correcting current on first working electrode shows remarkable disturbing effect in lower interferent concentration.This shows that the method for using formula 7a to proofread and correct the output current of first working electrode is being effective aspect the correction interference.Table 1 shows that for the interference of paracetamol, gentianic acid and uric acid, the current correction among the formula 7a is effective.Table 1 has also shown the normal concentration scope of the chaff interference of finding in blood.In addition, table 1 also shows, in 240mg/dL concentration of glucose level, and, the current correction among the formula 7a is effective.
Figure 13 is the exploded perspective illustration of test-strips 800, and test-strips 800 is skin layers that design lunges the user, so that physiological fluid is forced out and collects in the test-strips 800 with seamless way.Test-strips 800 comprises substrate 50, conductive layer 802, insulation course 804, reagent layer 820, bonding coat 830 and top layer 824.Test-strips 800 also comprises far-end 5 and near-end 60.
In test-strips 800, conductive layer 802 is first layers that are arranged on the substrate 50.As shown in figure 13, conductive layer 802 comprises second working electrode 806, first working electrode 808, contrast electrode 810, second contact point 812, first contact point 814, reference contact point 816, test-strips check-out console 17.Because the material of conductive layer 802 is identical with test-strips 800 with test-strips 62 with the method that is used to print conductive layer 802.
The blade 826 that also can be called penetrating parts can be suitable for thrusting user's skin and blood is drawn in the test-strips 800, and such second working electrode 806, first working electrode 808 and contrast electrode 810 are wetted.Blade 826 is included in the blade substrate 832 that stops on the distal portions 58 of test-strips of assembling.Blade 826 can by insulating material for example plastics, glass and silicon or conductive material for example stainless steel and gold are made.The further describing of integration medical treatment device of use integration blade can be referring to International Application PCT/GB01/05634 and U.S. patented claim 10/143,399.In addition, blade 826 can for example be made by the progressive die stamping technology, and described technology is as being disclosed in above-mentioned International Application PCT/GB01/05634 and the U.S. patented claim 10/143,399.
Figure 14 is the simplified schematic diagram illustrating that shows the measuring instrument 900 that is connected with test-strips.In one embodiment of the invention, following test-strips can be suitable for using together with measuring instrument 900: test-strips 62, test-strips 162, test-strips 800, test-strips 2000, test-strips 3000 or test-strips 5000.Measuring instrument 900 has at least 3 electrical pickofves, and these electrical pickofves formation are electrically connected with second working electrode, first working electrode and contrast electrode.Particularly, second contact point (13,103 or 812) is connected with first voltage source 910 with reference contact point (15,105 or 816); First contact point (11,101 or 814) is connected with second voltage source 920 with reference contact point (15,105 or 816).
When measuring, first voltage source 910 applies first current potential E1 between second working electrode and contrast electrode; Second voltage source 920 applies second current potential E2 between first working electrode and contrast electrode.In one embodiment of the invention, first current potential E1 and second current potential E2 can be identical, for example are about+0.4V.In another embodiment of the invention, first current potential E1 and second current potential E2 can be different.Apply blood sample, such second working electrode, first working electrode and contrast electrode are covered by blood.This allows the electric current that second working electrode and first working electrode measurement are directly proportional with glucose and/or non-enzyme particular source.Apply sample after 5 seconds, measuring instrument 900 is measured the oxidation current of second working electrode and first working electrode.
Table 1. uses the summary of the chaff interference influence of proofreading and correct and not proofreading and correct output current
| Pattern | Chaff interference | Concentration of glucose (mg/dL) | Influence is significant interferent concentration | The normal concentration scope of chaff interference |
| Do not proofread and correct | Paracetamol | 70 | 11 | 1-2 |
| Do not proofread and correct | Gentianic acid | 70 | 10 | 0.05-0.5 |
| Do not proofread and correct | Uric acid | 70 | 5 | 2.6-7.2 |
| Do not proofread and correct | Paracetamol | 240 | 16 | 1-2 |
| Do not proofread and correct | Gentianic acid | 240 | 12 | 0.05-0.5 |
| Do not proofread and correct | Uric acid | 240 | 8 | 2.6-7.2 |
| Proofread and correct | Paracetamol | 70 | 120 | 1-2 |
| Proofread and correct | Gentianic acid | 70 | 47 | 0.05-0.5 |
| Proofread and correct | Uric acid | 70 | 33 | 2.6-7.2 |
| Proofread and correct | Paracetamol | 240 | 59 | 1-2 |
| Proofread and correct | Gentianic acid | 240 | 178 | 0.05-0.5 |
| Proofread and correct | Uric acid | 240 | 29 | 2.6-7.2 |
Claims (2)
1. the method for the interference when measuring glucose in the reduction electrochemical sensor, described method comprises:
Measurement first electric current on first working electrode, described first working electrode is covered by reagent layer;
Second electric current of measurement on second working electrode, wherein said reagent layer partly covers described second working electrode, and described second working electrode has area coverage and area coverage not; With
Use following formula to calculate the correcting current density value of representing concentration of glucose:
Wherein G is the correcting current density value, WE
1Be the not correcting current density on described first working electrode, WE
2Be the not correcting current density on described second working electrode, A
CovBe the spreading area of described second working electrode, and A
UncBe the uncoated area of described second working electrode.
2. the method for the interference when measuring glucose in the reduction electrochemical sensor, described method comprises:
Measurement first electric current on first working electrode, wherein reagent layer partly covers described first working electrode, and described first working electrode has not area coverage of first area coverage and first;
Second electric current of measurement on second working electrode, wherein said reagent layer partly covers described second working electrode, and described second working electrode has not area coverage of second area coverage and second; With
Use following formula to calculate the correcting current density value of representing concentration of glucose:
Wherein
f
1＝A
cov1/A
unc1；
f
2＝A
cov2/A
unc2；
A
Unc1It is the uncoated area of described first working electrode;
A
Unc2It is the uncoated area of described second working electrode;
A
Cov1It is the spreading area of described first working electrode;
A
Cov2It is the spreading area of described second working electrode;
G is the correcting current density value;
WE
1Be the not correcting current density on described first working electrode, and
WE
2Be the not correcting current density on described second working electrode.
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| US60/558,424 | 2004-03-31 |
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| CN 200480039546 Pending CN1902481A (en) | 2003-10-31 | 2004-10-29 | Method of reducing the effect of direct and mediated interference current in an electrochemical test strip |
| CNB2004800395265A Expired - Fee Related CN100473982C (en) | 2003-10-31 | 2004-10-29 | Method for reducing interferences in an electrochemical sensor using two different applied potentials |
| CN200480039527XA Expired - Fee Related CN101163963B (en) | 2003-10-31 | 2004-10-29 | A measurer of reducing interferences in an electrochemical sensor using two different applied potentials |
| CN 200910002047 Expired - Fee Related CN101533007B (en) | 2003-10-31 | 2004-10-29 | Method of reducing the effect of direct interference current in an electrochemical test strip |
| CN 200910007080 Expired - Fee Related CN101493466B (en) | 2003-10-31 | 2004-10-29 | Method for reducing interference in electrochemical sensor using two different applied potentials |
| CN 200480039541 Pending CN1902479A (en) | 2003-10-31 | 2004-10-29 | Electrochemical test strip for reducing the effect of direct and mediated interference current |
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| CN200480039527XA Expired - Fee Related CN101163963B (en) | 2003-10-31 | 2004-10-29 | A measurer of reducing interferences in an electrochemical sensor using two different applied potentials |
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| CN104569102B (en) * | 2015-02-04 | 2018-04-06 | 苏州市玮琪生物科技有限公司 | Detect the bio-sensing electrode and method of Blood Trace signal |
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| JP6778058B2 (en) * | 2016-08-31 | 2020-10-28 | シスメックス株式会社 | Sensor assembly, test substance monitoring system and test substance monitoring method |
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| Publication number | Publication date |
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| CN101163963A (en) | 2008-04-16 |
| CN101493466B (en) | 2013-11-06 |
| CN1902481A (en) | 2007-01-24 |
| CN1902478A (en) | 2007-01-24 |
| CN100473982C (en) | 2009-04-01 |
| CN101533007A (en) | 2009-09-16 |
| CN1902477A (en) | 2007-01-24 |
| CN1902479A (en) | 2007-01-24 |
| CN101493466A (en) | 2009-07-29 |
| CN101533007B (en) | 2013-01-02 |
| CN101163963B (en) | 2011-05-04 |
| CN1902480A (en) | 2007-01-24 |
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