CN100372940C - Method for preparing lactate biosensing strip - Google Patents
Method for preparing lactate biosensing strip Download PDFInfo
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- CN100372940C CN100372940C CNB028301927A CN02830192A CN100372940C CN 100372940 C CN100372940 C CN 100372940C CN B028301927 A CNB028301927 A CN B028301927A CN 02830192 A CN02830192 A CN 02830192A CN 100372940 C CN100372940 C CN 100372940C
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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
- C12Q1/002—Electrode membranes
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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
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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
- C12Q1/005—Enzyme electrodes involving specific analytes or enzymes
Abstract
The present invention provides a manufacturing method for a lactate biosensing strip used for analyzing lactic acid in a sample. The sensor comprises a dry strip sensor of electric conducting material, and the dry strip sensor at least comprises an outer surface i, a silk-screen printing reference electrode ii and a silk-screen printing working electrode iii.
Description
Technical Field
The invention relates to a lactate biosensing strip for measuring lactate solution. The invention also relates to a method for manufacturing the novel lactate biosensing strip and the use thereof in lactate sensing.
Background
Depending on the individual examination and clinical laboratory results, a physician can determine the presence and concentration of biological analytes in a critically ill patient. In a well-controlled, high-quality environment, clinical laboratories can provide a variety of automated systems for a large number of tests and analytical supports. However, clinical laboratories are unable to provide the immediate results needed to adequately treat trauma and multi-organ dysfunction/failure patients. To meet the clinical need for immediate test results, several technologies are emerging that are being tested at the bedside using reliable, automated analytical instruments including electrochemical biosensors, optical fluorescence sensors, paramagnetic particles for coagulation detection systems, and micro-electromechanical devices for chemical and immunochemical detection. While rapid operation of chemical panels with multiple analytes is possible, these techniques also address a priori obstacles such as calibration of detection devices.
These tests can be classified as: 1) in vitro, it can be performed at the bedside; 2) in vitro (ex vivo) or para vivo, which can be performed at the wrist; and 3) in vivo, which may be performed in a patient. Such testing can provide indirect cost effectiveness, savings such as reduced labor costs, reduced blood identification and errors in transportation, and reduced patient complications. In hospital departments including intensive care units, operating rooms, emergency departments (ER), interventional departments, general patient care units, and outpatient surgery and ambulatory care units, extracorporeal or bedside equipment is commonly employed. Similar to clinical laboratories, in vitro diagnostictests provide a wide range of diagnostic tests. Extracorporeal diagnostic test systems are typically not connected online to the patient and require an operator to sample the blood.
The main diagnostic test categories in the diagnostic market include arterial blood gases, blood chemistry, blood glucose, blood clotting, drug abuse testing, hemoglobin, blood cell volume, infectious diseases, and therapeutic drug monitoring. Other categories include cancer markers, cardiac markers, cholesterol detection, immunodetection, infectious disease detection, lactate, and thrombolytic monitoring.
In vitro diagnostics use external sensors for online real-time detection with little to no blood loss. In general, the sampled blood flows through a closed system to reduce blood contact. In vitro systems can minimize problems associated with in vivo sensors in patients including clotting, inaccuracies, calibration drift, and the inability to recalibrate immediately. An exemplary ex vivo system is disclosed in U.S. patent application No. 5,505,828.
In vivo diagnostics has considerable potential in the treatment of patients who are extremely critical and unstable. Despite the ongoing development of many companies, the technical barriers to in vivo sensors have so far kept them away from general commercial applications.
Because in vitro and in vivo diagnostics are on-line systems, they can reduce quality control and information integration errors that occur in clinical or in vitro testing. Quality control errors are typically due to operator error, rather than instrument error or equipment failure. Examples of errors include improper sample volume, inaccurate calibration, use of failed test strips, inadequate validation, improper instrument maintenance, incorrect timing of the test procedure, and use of wrong materials. Clinical information system integration enablestest data collected at the bedside to be entered directly into the patient's records. Such an approach increases the efficiency of the patient management process, integrates the laboratory information system and the clinical information system, and provides a "seamless" flow of all kinds of patient information.
Lactic acid is a by-product of sugar metabolism, and the product of glycolysis (pyruvate) is converted to lactic acid in an anaerobic environment, i.e., in the absence of oxygen in the cell. Estimation of lactic acid content is therefore of great importance in respiratory disorders, heart diseases, reduced labour and the like. The concentration of lactic acid in the blood of normal human is in the range of 1.2-2.7 mM.
For example, the method of lactate determination employs a variety of chemical and physical techniques. Traditional assays involve chemical treatment of lactate in human blood and converting it to a chromogenic product that can be measured spectrophotometrically, which involves reacting the blood with an enzyme known as Lactate Dehydrogenase (LDH) in the assay. Since NADH is formed in such a process, the absorbance at 340nm is detected, which is a method for detecting lactic acid originally present in blood.
Us patent No. 6,117,920 discloses an on-line lactate sensor device. The sensor device includes a lactate sensor, a conduit for aspirating a test sample, and a first liquid flow line providing fluid communication between the lactate sensor and the conduit. The sensor device also includes a source of sensor calibration and anti-clotting solution and a second fluid flow line providing fluid communication between the source of sensor calibration and anti-clotting solution and the lactate sensor.
In practice, there are difficulties in applying such a detection method to the detection of blood samples. Disadvantages of this approach include lack of specificity, difficulty of standardization, requirement of large amounts of blood, and use of unstable and corrosive reagents. Such methods also involve optical detection and are therefore expensive and time consuming. But the samples also need to be prepared. Another disadvantage is that the detection of lactate levels using the above-mentioned prior art methods needs to be done in the laboratory by qualified personnel.
Asha Chaubey et al, in Electrochimica Acta Vol 46,723-729(2000), disclose the curing of lactate dehydrogenase on electrochemically treated polypyrrole polyvinylsulfonate synthetic films. The reaction time is reported to be about 40 seconds and the pot life under refrigerated conditions is about 2 weeks. In another disclosure (Asha Chaubey et al, Analyticla Chimica Acta Vol49, 98-103, 2000), the curing of lactate dehydrogenase on conductive polyaniline films is disclosed. The reaction linear concentration is 0.1mM to 1mM lactic acid concentration, and the shelf life under refrigerated conditions is about 3 weeks. It is therefore desirable to obtain sensors with longer lifetimes and shorter reaction times.
Accordingly, it is important to provide a lactate biosensing strip that overcomes the above-mentioned drawbacks without losing detection efficiency and accuracy.
Object of the Invention
The main object of the present invention is to provide a novel lactate biosensing strip for the detection of lactate in an aqueous medium.
It is another object of the present invention to provide a lactate biosensing strip that quickly and accurately assesses lactate in an aqueous medium.
It is a further object of the present invention to provide a lactate biosensing strip that is low cost and can even be used bynon-medical personnel.
It is a further object of the present invention to provide an assay which can be performed without elaborate pre-treatment of the blood sample.
It is another object of the present invention to provide a lactate biosensing strip having a high activity of 75%.
It is another object of the present invention to provide a lactate sensor strip that can read results in situ.
Summary of The Invention
Compared with the conventional method, the lactate biosensing strip has many advantages such as rapid reaction, small size convenience, specific reaction, no need of any sample preparation, low cost and high sensitivity of detection. Compared with the traditional method, the sensor has the greatest advantage that the sample operation can be completed by ordinary people.
The present invention provides a lactate biosensing strip for analyzing lactate in a sample, said sensor comprising a dry strip sensor of electrically conductive material having at least:
i. an outer surface of the outer shell,
a screen printed reference electrode, and
a screen printed working electrode.
Accordingly, the present invention provides a lactate sensor comprising an electrically insulating base support (1); a pair of spaced apart first and second silver layers (2) deposited on the surfaces thereof; a pair of graphite layers, each graphite layer of said pair being deposited on and electrically connected to a respective silver layer (2), said first silver layer being completely covered by a respective graphite layer, said second silver layer being partially covered in its middle by a respective graphite layer, leaving a connecting and working area where said layers are uncovered; an Ag/AgCl electrode (4) on the surface of the working area of the second silver layer electrode layer; lactate oxidase deposited on the graphite layer working area covering the first silver layer together with the medium; the silver/silver chloride reference electrode (4) and a working electrode of enzyme and medium are loaded on the support (1); the entire working area of the reference electrode and the working electrode is covered with a hydrophilic membrane.
In one embodiment of the invention, the electrically insulating base support used is made of polyvinyl chloride.
In one embodiment of the invention, the distance between the silver layers is between 0.5 and 1 mm.
In another embodiment of the present invention, the thickness of each silver layer is between 15 and 25 microns.
In another embodiment of the present invention, the electron medium layer comprises a potassium ferricyanide layer or a ferrocene layer.
In another embodiment of the present invention, the hydrophilic membrane is made of nylon or polyester.
In another embodiment of the invention, the working area of the electrode is a target area for dispensing an analyte sample.
In another embodiment of the invention, the connection end region of the electrode is a region for connecting the electrode to an electrometer (electrometer).
The lactate biosensing strip of the present invention has 75% activity and 30-40 seconds lactate detection reaction time. The shelf life of the lactate biosensing strip of the present invention is about 4 months under refrigerated conditions. The life span of the biosensing strip is about 2 months at room temperature (25-30 ℃). The sensor strip ofthe present invention is disposable.
The invention also relates to a method for manufacturing a lactate biosensing strip, said strip comprising an electrically insulating base support (1); a pair of spaced-apart first and second silver layers (2) deposited thereon; a pair of graphite layers, each graphite layer of said pair of graphite layers being deposited on and electrically connected to a respective silver layer (2); the first silver layers are completely covered by respective graphite layers; the second silver layer is partially covered in the middle by a corresponding graphite layer, leaving a connecting and working zone uncovered by said layer; an Ag/AgCl electrode (4) provided on a surface of the working area of the second silver layer electrode layer; lactate oxidase (5) deposited with media on the working area of the graphite layer covered with the first silver layer; a silver/silver chloride electrode (4) and a working electrode of enzyme and medium are loaded on the support (1); the method comprises the following steps of,
(a) depositing a pair of silver layers on an electrically insulating base support by any conventional method;
(b) depositing a pair of graphite layers on said silver layers by any conventional method, each of said silver layers being deposited by a graphite layer, said first graphite layer completely covering said first silver layer on the side of said silver layer facing away from the surface of said base support, and said second graphite layer partially covering said second silver layer;
(c) depositing a silver chloride layer on the portion of the second silver layer where the graphite layer is not deposited to obtain a silver/silver chloride electrode;
(d) physically adsorbing lactate oxidase through an electronic medium on the first graphite layer deposited on the silver layer to obtain a workingelectrode; and
(e) the application of an outer hydrophilic membrane on the first reference electrode and the second working electrode described above results in a single-component desired electrode pair on an electrically insulating base support.
In one embodiment of the invention, the electrically insulating base support comprises polyvinyl chloride.
In another embodiment of the invention, the silver layer used is applied by a screen printing step.
In another embodiment of the invention, the graphite layer used is applied by a screen printing step.
In another embodiment of the present invention, the sample to be measured is an aqueous solution of lactic acid or a blood sample having a volume of 25 to 30. mu.l.
In another embodiment of the invention, the electron mediator used is selected from potassium ferricyanide and ferrocene.
In another embodiment of the invention, the connecting electrode tip region is a region for connecting the electrode to an electricity meter.
In another embodiment of the present invention, the hydrophilic membrane is made of nylon or polyester.
Brief description of the drawings
FIG. 1 is a schematic view of a biosensor strip according to the present invention.
FIG. 2 shows the reaction curve of the lactate biosensing strip of the present invention against a standard lactate test sample.
Fig. 3 shows a sensor calibration curve for a standard lactate test sample prepared in the laboratory.
FIG. 4 shows the shelflife stability characteristics of the lactate strips of the present invention.
Detailed description of the invention
As shown in fig. 1, the present invention includes an electrically insulating base support (1) supporting electrode assemblies (2), (3), (4) and (5). The electrode assembly comprises two electrode systems: working electrode systems (2), (3) and (5) consisting of a layer of silver on which a layer of graphite is deposited, and a layer of enzymes and media absorbed in an inorganic matrix. The other electrode assembly includes a reference electrode formed of a silver layer having a graphite layer and a silver/silver chloride layer partially deposited thereon. FIG. 1 shows a PVC substrate (i) comprising an electrode support substrate. Conductive silver tracks (ii) a layer of conductive graphite (iii) is screen printed on the surface of the conductive silver tracks to connect the sensor to a reading device.
The target area consists of the working electrode (iv) and a reference electrode (v) applied to the end of the rail by screen printing. Coating an insulating layer on the printed electrodes to protect the printed electrodes; the main portion may be coated with one or more descriptions. The conductive graphite track (ii) does not extend the full length of the reference electrode and the silver track.
To achieve calibration of the biosensing strip, the strip was used to detect the current in solution when the lactate concentration was between 1-8 mM. The detection current for each concentration was measured and plotted as shown in fig. 2. In FIG. 2, curve (1) is a reaction curve corresponding to a 1mM lactic acid solution, curve (2) is a 2mM solution, curve (3) is a 4mM solution, curve (4) is a 6mM solution, and curve (5) is a 5mM solution. This indicates that the biosensing strip of the present invention can be used to detect lactate in a blood sample if the concentration in the individual is in the region of 1-8 mM. By analyzing the time variation of the strip current, the sensitivity of the system with respect to the reaction time required to obtain a stable current value can be determined. This includes the time from the time a drop of standard test solution is placed on the strip for current testing until the current asymptotically reaches a stable value. It was observed (fig. 3) that the current reached a steady value in 30-40 seconds. The expiration date profile was determined by measuring the current produced by a known concentration of lactic acid on a sensor strip stored for various periods of time. The data are shown in figure 4. In FIG. 4, curve (1) corresponds to a sensor strip stored at refrigerated conditions (4 ℃) and curve (2) corresponds to a sensor strip stored at 25-30 ℃.
The present invention provides a method of making a lactate sensor strip comprising forming first and second electrodes on a substrate by coating a silver layer for each of said electrodes; applying a graphite layer to silver chloride in the operative region of the second electrode; the mediator and enzyme are coated on the graphite layer of the working area of the first electrode. An outer hydrophilic membrane is applied in the region of the first electrode. The silver and graphite layers are preferably applied by a screen printing step.
The main feature of the invention is that the sensor is a dry strip sensor. It was found that similar mixing of reagents in a wet sensor system did not produce good results in the expected range of detectable lactate concentrations. The invention comprises a substrate supporting an electrode assembly comprising two electrode systems, a working electrode supported by the substrate and deposited in spaced relation to one another and another electrode serving as a reference electrode. The lactate sensor strip includes a substrate carrying a first or working electrode and a second or reference electrode, the electrodes being deposited in a spaced relationship to each other. The first electrode is a working electrode and has an end section that extends into the working region through the operating region. The second electrode is a reference electrode and has an end section that extends through the operating region into the working region. In both cases, the material of the respective ends is different for the base conductive layers of the first and second electrodes.
Commercially available lactate oxidase was mixed in phosphate buffer and the appropriate amount of this solution was then injected onto the pre-printed working electrode. The solution is dried at a permissible temperature and subsequently
i. The printing of the electrically conductive tracks is carried out,
printing of a reference electrode,
printing of the working electrode(s),
fixation of the membrane on the electrode.
Along the operating and working areas, the working electrode and the reference electrode each comprise a silver material base conductive layer. A graphite layer is deposited on the silver layer of the working electrode and extends to the end; the graphite layer is applied to the operating region of the reference electrode and extends to the terminal end. Ag/AgCl is deposited on the target area of the reference electrode. The working electrode comprises a conductive surface with a mediator complex and lactate oxidase. When this catalytic activity occurs, the mediator compound transfers the electrode from the enzyme to the electrode. A hydrophilic membrane must be provided on the working area of the electrode. The surfactant can break down lipoprotein complexesin the blood, and lactic acid is subsequently oxidized to pyruvic acid by lactate oxidase. The mediator compound is electrochemically reduced at the electrode to produce a current that is detectable at the electrode, which current is correlated with the activity of lactic acid oxidation, thereby deriving the amount of lactic acid present in the sample, which current is produced by a series of coupling reactions:
l-lactic acid + LOD(Oxidation)-------- - - -pyruvic acid + LOD(reduction)
LOD(reduction)+Me(Oxidation)------LOD(Oxidation)+Me(reduction)
The redox mediator is oxidized at the base electrode and generates a current proportional to the concentration of lactic acid. The current may be measured by any conventional electronic system.
The following examples are given by way of illustration only and should not be construed to limit the scope of the present invention.
Example 1: preparation of graphite paste containing Medium
A screen-printable working electrode graphite paste was prepared by mixing 100mg of graphite powder and polyvinylpyrroline (binder) with 0.01M potassium ferricyanide (medium) in ethylene glycol monobutyl ether.
Example 2: preparation of dried bars
A commercially available lactate oxidase solution (2. mu.l) containing 2U lactate oxidase was physically absorbed on a sensor strip mixed with a graphite electrode medium and kept at 25 ℃ overnight for drying. And coating a layer of hydrophilic nylon membrane on the dried electrode. Before applying the membrane, it was put in distilled water containing 10% surfactant (Tween 80) for a certain period of time, and then the dried membrane was fixedon the strip.
Example 3: preparation of lactic acid Standard lactic acid solution
A10 mM stock lactic acid solution was prepared in 0.1M phosphate buffer. The stock solutions were diluted with phosphate buffer to give standard solutions of 2mM, 4mM, 6mM and 8 mM.
Example 4: preparation of enzyme stock solutions
15mg of lactate oxidase was dissolved in 100. mu.l of 0.1M phosphate buffer to obtain a stock solution with a concentration of 5U/. mu.l, and the stock solution was further diluted to 1U/. mu.l to obtain an enzyme working solution.
Example 5: curing of enzymes on media-mixed graphite drying strips
Mu.l of enzyme solution containing 2U lactate oxidase was physically absorbed on a media-mixed graphite electrode strip and kept at 25 ℃ overnight for drying. The dried electrode was coated with a hydrophilic nylon membrane. Before the application of the membranes, they were placed in distilled water containing 10% surfactant (Tween 80) for a period of time, and the dried membranes were subsequently fixed to the strips.
Example 6: enzyme activity
Lactate oxidase activity was estimated using the lactate oxidase activity protocol from Sigma. The basic principle is that lactate oxidase can convert L-lactic acid into pyruvic acid and H2O2。H2O2Subsequently converted by peroxidase into a chromogenic dye in the presence of 4-aminoantipyrine (4AAP) and Dimethylaniline (DMA).
L-lactic acid + O2→ pyruvic acid + H2O2
2H2O2+4-AAP + DMA → quinonediimine dye + H2O
Under the optimal conditions of 37 ℃ and 6.5 pH, the dye can generate absorption at the light path of 1cm and 565 nm.
The activity of the immobilized enzyme was calculated by the following formula:
Ucm-2＝AV/∈ts
wherein A is the change in absorbance before and after incubation
V is the total volume (3ml)
E is the millimolar extinction coefficient at 565nm for quinone diimine dye (35.33)
t is the reaction time (10 minutes)
s is the surface area of the enzyme electrode
The enzyme activity of the LOD immobilized on the working graphite strip was found to be 75%.
Example 7: investigation of amperometric response
The lactate biosensing strip, which contained an enzyme (LOD) immobilized on the graphite surface as the working electrode and an Ag/AgCl reference electrode connected to the input of an electrometer, was polarized at a bias voltage of 0.4V to perform the measurement of the amperometric calibration reaction of lactate (1-8mM) (fig. 2). The maximum current of 60. mu.A was obtained in 8mM lactic acid solution, and no significant change in current was observed even at higher concentrations. The reaction time of the lactic acid solution (1-8mM) was 40 seconds for each concentration of lactic acid (FIG. 3). The results have a reproducibility within 5%. The following principles relate to amperometric measurements:
lactic acid + LOD(Oxidation)→ pyruvic acid + LOD(reduction)
LOD(reduction)+Fe3+→LOD(Oxidation)+Fe2+
Advantages of the invention
1. The lactate biosensing strip allows for rapid assessment of lactate in a sample.
2. The samples were shelf-stable for up to 4 months under refrigerated conditions.
3. At lactic acid concentrations of 1-8mM, the strip had a linear response.
4. The strip is disposable and does not pose any environmental hazard.
5. The strip can be easily used even by persons without formal medical training.
Claims (10)
1. A method of making a lactate biosensing strip, said biosensing strip comprising an electrically insulating base support; a pair of spaced apart first and second silver layers deposited on the surfaces thereof; a pair of graphite layers, each graphite layer of said pair being deposited on and electrically connected to a respective silver layer, said first silver layer being completely covered by a corresponding graphite layer, leaving uncovered terminal and working regions, said second silver layer being partially covered by a corresponding graphite layer in the middle thereof; an Ag/AgCl electrode provided on a surface of the working region of the second silver electrode layer; lactate oxidase deposited on the graphite layer working area covering the first silver layer together with the medium, the silver/silver chloride electrode and the enzyme layer working electrode containing the medium are carried on the support, the working areas of the silver/silver chloride reference electrode and the working electrode are covered with a hydrophilic membrane,
the lactate biosensing strip exhibiting 75% activity and a lactate detection reaction time in the range of 30-40 seconds, a shelf life of about 4 months under refrigerated conditions and about 2 months at room temperature, the strip being disposable, the method comprising,
(a) depositing a pair of silver layers on an electrically insulating base support;
(b) depositing a pair of graphite layers on said silver layers, each of said silver layers being deposited with a graphite layer, the first graphite layer completely covering the first silver layer and the second graphite layer only partially covering the second silver layer on its back side facing said base support;
(c) depositing a silver chloride layer on the part of the second silver layer on which the graphite layer is not deposited to obtain a silver/silver chloride electrode;
(d) physically adsorbing lactate oxidase through an electronic medium on a first graphite layer deposited on the silver layer to obtain a working electrode; and
(e) the application of an outer hydrophilic membrane on the first reference electrode and on the second working electrode described above results in the desired pair of electrodes in the form of a single component on an electrically insulating base support.
2. The method of claim 1, wherein the electrically insulating base support used is polyvinyl chloride.
3. The method of claim 1, wherein the silver layer is applied by a screen printing step.
4. The method of claim 1, wherein the graphite layer is applied by a screen printing step.
5. The method of claim 1, wherein the test sample is a 25-30 μ l volume of aqueous lactic acid solution or blood sample.
6. A process as claimed in claim 1, wherein the electron mediator used is selected from potassium ferricyanide and ferrocene.
7. The method of claim 1, wherein the working area of the electrode is a target area for dispensing an analyte sample.
8. The method of claim 1, wherein the connection terminal region of the electrode is a region for connecting the electrode to an electricity meter.
9. The method of claim 1, wherein the hydrophilic membrane is made of nylon or polyester.
10. The method of claim 1, wherein the room temperature conditions are 25-30 ℃.
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/IB2002/005677 WO2004058992A1 (en) | 2002-12-31 | 2002-12-31 | Method for preparing lactate biosensing strip |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1742092A CN1742092A (en) | 2006-03-01 |
| CN100372940C true CN100372940C (en) | 2008-03-05 |
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|---|---|---|---|
| CNB028301927A Expired - Fee Related CN100372940C (en) | 2002-12-31 | 2002-12-31 | Method for preparing lactate biosensing strip |
Country Status (6)
| Country | Link |
|---|---|
| EP (1) | EP1578984A1 (en) |
| JP (1) | JP2006512571A (en) |
| CN (1) | CN100372940C (en) |
| AU (1) | AU2002368512A1 (en) |
| CA (1) | CA2512279C (en) |
| WO (1) | WO2004058992A1 (en) |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| GB0606998D0 (en) * | 2006-04-06 | 2006-05-17 | Oxford Biosensors Ltd | Lipoprotein sensor |
| CN101344501B (en) * | 2008-08-22 | 2011-10-05 | 中南大学 | Silk screen printing electrode, manufacturing technique and use thereof |
| JP2010230369A (en) * | 2009-03-26 | 2010-10-14 | Ryukoku Univ | Electrode structure, manufacturing method of the same, and electrochemical sensor |
| CN101650331B (en) * | 2009-08-27 | 2012-09-05 | 无锡爱康生物科技有限公司 | Enzyme bioelectrochemical sensing chip and preparation method thereof |
| KR101947639B1 (en) * | 2017-08-08 | 2019-02-14 | 한국화학연구원 | Glucose sensor for ex-vivo diagnosis comprising graphene-based electrode |
| US20190320947A1 (en) * | 2018-04-19 | 2019-10-24 | Abbott Diabetes Care Inc. | Lactate sensors and associated methods |
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| US5707502A (en) * | 1996-07-12 | 1998-01-13 | Chiron Diagnostics Corporation | Sensors for measuring analyte concentrations and methods of making same |
| US5820551A (en) * | 1983-05-05 | 1998-10-13 | Hill; Hugh Allen Oliver | Strip electrode with screen printing |
| WO1999019507A1 (en) * | 1997-10-16 | 1999-04-22 | Abbott Laboratories | Biosensor electrode mediators for regeneration of cofactors |
-
2002
- 2002-12-31 WO PCT/IB2002/005677 patent/WO2004058992A1/en active Application Filing
- 2002-12-31 CN CNB028301927A patent/CN100372940C/en not_active Expired - Fee Related
- 2002-12-31 CA CA2512279A patent/CA2512279C/en not_active Expired - Fee Related
- 2002-12-31 AU AU2002368512A patent/AU2002368512A1/en not_active Abandoned
- 2002-12-31 EP EP02790636A patent/EP1578984A1/en not_active Withdrawn
- 2002-12-31 JP JP2004563409A patent/JP2006512571A/en active Pending
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5820551A (en) * | 1983-05-05 | 1998-10-13 | Hill; Hugh Allen Oliver | Strip electrode with screen printing |
| US5707502A (en) * | 1996-07-12 | 1998-01-13 | Chiron Diagnostics Corporation | Sensors for measuring analyte concentrations and methods of making same |
| WO1999019507A1 (en) * | 1997-10-16 | 1999-04-22 | Abbott Laboratories | Biosensor electrode mediators for regeneration of cofactors |
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| JP2006512571A (en) | 2006-04-13 |
| CA2512279A1 (en) | 2004-07-15 |
| WO2004058992A1 (en) | 2004-07-15 |
| CA2512279C (en) | 2010-06-29 |
| CN1742092A (en) | 2006-03-01 |
| EP1578984A1 (en) | 2005-09-28 |
| AU2002368512A1 (en) | 2004-07-22 |
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