WO2002093153A1 - Cellule electrochimique de mesure de debit - Google Patents

Cellule electrochimique de mesure de debit Download PDF

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Publication number
WO2002093153A1
WO2002093153A1 PCT/EP2002/004681 EP0204681W WO02093153A1 WO 2002093153 A1 WO2002093153 A1 WO 2002093153A1 EP 0204681 W EP0204681 W EP 0204681W WO 02093153 A1 WO02093153 A1 WO 02093153A1
Authority
WO
WIPO (PCT)
Prior art keywords
measuring
containment
substrate
measuring cell
chamber
Prior art date
Application number
PCT/EP2002/004681
Other languages
German (de)
English (en)
Inventor
Carlo Stefan Effenhauser
Holger Kotzan
Reinhard Kotulla
Michael Hein
Original Assignee
Roche Diagniostics Gmbh
F.Hoffman-La Roche Ag
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Roche Diagniostics Gmbh, F.Hoffman-La Roche Ag filed Critical Roche Diagniostics Gmbh
Publication of WO2002093153A1 publication Critical patent/WO2002093153A1/fr

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/403—Cells and electrode assemblies

Definitions

  • the invention relates to an electrochemical flow measuring cell for performing an electrochemical or biosensor measurement on an analyte.
  • a measurement can be, for example, the acquisition, i.e. the qualitative and / or quantitative determination of an analytical parameter of the analyte, e.g. the concentration of a chemical or biochemical component of the analyte.
  • Electrochemical measuring cells comprise a measuring chamber into which an analyte to be analyzed can be introduced.
  • the measuring chamber serves as a reservoir for the analyte to be analyzed.
  • the analyte is introduced into the measuring chamber for a discontinuous or one-off measurement (so-called
  • the analyte can also be fed to the measuring chamber by transport in a transport channel.
  • the measuring chamber is referred to as a flow chamber or flow measuring chamber since the analyte is passed through the measuring chamber.
  • the analyte is supplied to the measuring chamber or flow chamber in With such a flow measuring cell, the continuous acquisition of analytical parameters is possible continuously.
  • the flow chamber is often referred to as a channel or micro-channel system.
  • Such measuring cells also include a containment, which is also referred to as the actual measuring cell. It has a substance-recognizing component, e.g. an enzyme or an ionophore, can be filled to determine an analytical parameter of the analyte and is connected to the measuring chamber.
  • substance-recognizing components are sometimes referred to as membrane material if they are membrane-bound.
  • Miniaturized measuring cells with integrated chemical and / or biosensor elements are known. They include ion-selective electrodes that contain an electroactive substance (ionophore) that determines the ion selectivity of the sensor element.
  • the electroactive substance i.e. the substance-recognizing component is often bound in the form of a membrane or a gel.
  • the areas of application of electrochemical measuring cells are very diverse.
  • Known examples are the integration into flow systems, for example flow injection analysis, and microdialysis systems for determining the glucose concentration in human tissue.
  • Containment principle is produced and works with appropriately doped polymer membranes and gel layers for substance detection.
  • the polymer membranes and gel layers are not applied as a thin layer on the surface of silicon chips, but rather as a "bulk" volume implemented in the chip. This has considerable advantages over planar structures in terms of simple manufacture and high long-term stability.
  • Such sensors in silicon technology can also be installed in prefabricated measuring cells.
  • the miniaturized chemical and biosensor known from WO 92/21020 consists of a substrate which is designed as a plate-shaped carrier in which a containment is introduced. It has an opening on each side of the carrier, the containment tapering from one side of the carrier to the other. The analyte flows directly past the smaller opening of the containment.
  • the document WO 00/62931 AI relates to a measuring cell for spot measurements.
  • the analyte is mixed with a reagent and transferred to the containment, in which an electrochemical measurement takes place.
  • the analyte is thus completely transferred into the containment or the containment is completely filled with the analyte.
  • a complete "bleeding" of the measuring cell takes place.
  • Such a measuring cell can therefore only be used once for a single measurement or has to be regenerated in a complex manner.
  • the regeneration requires a large number of process steps, namely rinsing with rinsing solutions, applying new reagent solutions, calibrating the measuring cell and additional equipment on valves, pumps, channel systems, etc., in order to carry out the regeneration with different liquids. Overall, such a regeneration is volume, time and cost consuming and does not result in the result that a continuous measurement of the analyzed analyte can be carried out.
  • the document WO 99/45382 AI relates to a distant prior art. It is not aimed at an electrochemical measuring cell, but at urine analysis test strips which are inserted into a device for the optical detection of particulate components. With such a measurement, no continuous measurement is possible, but only a one-time measurement.
  • an electrochemical flow measuring cell which works according to the containment principle and in which the flow chamber with the containment is integrated on a chip.
  • the flow chamber is connected to the containment via a small measuring opening in the containment.
  • the analyte is connected to the active sensor surface, which is wholly or partly formed by the electrodes on the walls of the containment.
  • the containment is completely penetrating the substrate; the substrate in the form of a plate-shaped carrier has opposite different sized openings between which the containment extends.
  • the smaller opening of the containment is covered by a plate.
  • Flow chamber is formed in the substrate containing the containment, in the cover plate or in both, and the analyte flows directly over the smaller opening of the containment.
  • the filled containment structure is covered with an encapsulation layer.
  • Measuring cells which comprise a substrate in which recesses for the measuring chamber, for the containment and for a measuring opening connecting the measuring chamber with the containment are formed on structured surfaces, and a cover element which is connected to the substrate on the structured substrate surface and the recesses covers the measuring chamber of the containment and the measuring opening, should meet different requirements.
  • ion-selective sensor elements which, for example, have liquid membranes or other membranes or electrochemically or biochemically relevant sensor element coatings which are manufactured, for example, from a liquid phase or are equipped with solid-state membranes
  • this has the following properties: good and long-term stable membrane adhesion, minimal depletion Ionophores or other substance-recognizing reagents (enzymes) and auxiliary reagents in the membrane, an optimal condition for contacting and encapsulation and, above all, an inexpensive and simple manufacture of the measuring cell.
  • a major disadvantage of the measuring cells according to the prior art is furthermore that the known methods, particularly when using inexpensive plastic substrates, require a relatively high production outlay and cannot optimally utilize the possibilities of microstructuring.
  • the known methods either require a separate structuring of the front and back of a substrate plate with the associated technical problems, for example due to the required protection of the already structured side, which is not easy in particular in the case of etching processes, and due to the limited accuracy of the mutual alignment of the structures on the front opposite the back.
  • a separate structuring of two different layers is required, which must then be assembled with high precision, which places considerable positioning requirements in view of the small dimensions of the functionally relevant components.
  • plastic structures are to be realized, according to the state of the art, for example an injection molding process comes into question, which, due to the shrinking of the material after the molding process, leads to problematic shape changes which affect all further geometrically precise processing steps (for example vapor deposition of metal electrodes) a mask) has a negative effect.
  • the production of small measuring openings in silicon by etching on both sides is also problematic since the size of the opening is critically influenced by the etching time, which, however, can only be adhered to with great difficulty with great difficulty.
  • the object of the invention is to create electrochemical flow-through cells which can be manufactured in a simple, inexpensive and highly precise manner while avoiding considerable adjustment problems in a geometrically varied form of containment, a continuous one
  • An electrochemical flow measuring cell for carrying out an electrochemical or biosensory measurement on an analyte thus comprises a measuring chamber which can be filled with the analyte and which is designed as a flow measuring chamber, and a containment which is connected to the measuring chamber and which has a substance recognizer Component for determining an analytical parameter of the analyte can be filled.
  • the measuring cell comprises a substrate in which recesses for the measuring chamber, for the containment and for a measuring opening connecting the measuring chamber with the containment are formed on a structured substrate surface, and a cover element which is connected to the substrate on the structured substrate surface and covers the wells of the measuring chamber, the containment and the measuring opening.
  • An electrochemical flow measuring cell has the special feature that the depressions, which form the measuring chamber, the containment and the measuring opening, are formed on the structured substrate surface and do not penetrate the substrate in the direction of its thickness, so that the cavities formed by the depressions on it are surrounded by the material of the substrate and the cross-sectional area of the measuring opening in the substrate connecting the measuring chamber to the containment is designed as a constriction, so that the depletion of the substance-recognizing component from the containment when the analyte flows through the Flow measuring chamber is reduced. It has the advantages that it can be operated continuously and over a longer period of time without the need for regeneration.
  • An electrochemical flow measuring cell has the significant advantage from the point of view of manufacturing costs and manufacturing accuracy that the containment and the measuring chamber can be manufactured in a single operation on the same side of the substrate. This significantly simplifies production and solves the problem of adjusting the measuring chamber to the measuring opening of the containment, since all of them are for the Function of the measuring cell essential recesses can either be produced jointly and in parallel in one work step or can be produced in a sequential process on the same substrate side in the case of machining microfabrication or laser ablation.
  • Plastic substrates can be used, which can be processed with laser ablation structuring, which reduces the problem of shrinkage when cooling. This enables the subsequent application of metal layers with high accuracy using mask processes to implement the required feed and discharge lines.
  • the substance-recognizing component is mechanically anchored in the containment.
  • the containment measuring cell described represents, in the electrochemical sense, a so-called half-cell which is connected to at least one further electrochemical half-cell via an ion conductor (for example the analyte solution itself, an electrolyte bridge or a solid ion conductor) in order to implement a measurable unit.
  • an ion conductor for example the analyte solution itself, an electrolyte bridge or a solid ion conductor
  • a further half cell usually referred to as a reference electrode
  • Another half cell can be produced, for example, using the same manufacturing process as a containment cell.
  • this half cell is possible by a large number of methods known in the art, such as, for example, by sputtering, vapor deposition and / or electroplating of substrate surfaces or also by introducing solid metal electrodes in the form of wires, sheets or the like.
  • the containment and the measuring chamber are realized on a common substrate, which enables a monolithic integration of chemo and biosensors into microsystems.
  • Electrochemical measuring cells according to the invention can be produced individually or preferably also in large numbers on a substrate, in which case the measuring cells can be separated after their production.
  • a measuring cell according to the invention can be part of a microfluidic system, which can also contain other system components such as pumps, reaction lines or valves, which can be produced, for example, using known microstructure technologies.
  • the measuring chamber is designed as a flow-through chamber through which the analyte can be passed.
  • the measuring cell is therefore referred to as a flow measuring cell.
  • the measuring cells can advantageously be arranged linearly or in an array.
  • the manufacturing method according to the invention allows in a simple manner to produce multi-containment arrays in which more than one containment measuring cell in linear or two-dimensional configurations on the structured side of the sub- strat material are arranged. These arrays then enable the determination of N analytical parameters from M analytes, where N and M represent integers greater than or equal to 1.
  • N and M represent integers greater than or equal to 1.
  • An example of this would be the linear arrangement of containments on a flow channel, which contains several individual flow measuring chambers, for the determination of several analytical parameters from one analyte or also for the redundant detection of an analytical parameter with several identical measuring cells.
  • the invention achieves goals that the professional community has long sought.
  • FIG. 1 shows a plan view of the structured substrate surface of a first measuring cell according to the invention
  • FIG. 2 shows a section A-A with respect to FIG
  • FIG. 3 a section B-B 'to FIG. 1 with the cover element attached, FIG.
  • FIG. 4 shows a top view of the structured substrate surface of a second measuring cell according to the invention
  • FIG. 5 shows a section AA 1 to FIG. 4 with a cover element attached.
  • FIG. 1 shows a plan view of the structured substrate surface 1 of a measuring cell 2 according to the invention.
  • the substrate 3 there are depressions for a containment 4, a Measuring chamber 5 and a measuring opening 6 connecting the containment 4 to the measuring chamber 5 are introduced.
  • the measuring chamber 5 is designed as a channel-shaped flow measuring chamber and is flowed through by an analyte to be analyzed in the flow direction shown by the arrows.
  • the measuring cell 2 is therefore a flow measuring cell, with which a continuous detection of an analytical parameter of the analyte supplied through the measuring chamber 5 to the measuring cell 2 is possible.
  • the containment 4 has a circular cross section and is filled with a substance-recognizing component 7, also referred to as membrane material.
  • a substance-recognizing component 7 also referred to as membrane material.
  • Such a measuring cell 2 can be used, for example, to determine substances such as glucose, penicillin, urea, etc. in an analyte. So that the analyte in the measuring chamber 5 can come into contact with the substance-recognizing component 7 contained in the containment 4, the measuring chamber 5 is connected to the containment 4 via a measuring opening 6.
  • the cross-sectional area of the measuring opening 6 connecting the measuring chamber 5 to the containment 4 is formed in the substrate 3 as a narrowing of the containment 4.
  • Crosspieces 8 can serve to mechanically stabilize the arrangement, as an aid when filling the containment or to further narrow the measuring opening 6.
  • the depressions in the substrate 3 are preferably manufactured by means of a micromechanical manufacturing method.
  • a micromechanical manufacturing method for this purpose, depending on the material of the substrate 3, for example photolithographic processes, laser ablation, hot stamping, injection molding or machining mechanical microfabrication such as e.g. Micro-milling.
  • the substrate 3 preferably consists of a plastic, for example of PMMA or PC, which enable inexpensive production.
  • the measuring chamber 5 which is channel-shaped in the exemplary embodiment, is arranged on the structured substrate surface 1 next to the containment 4 and the measuring chamber 5 via the measuring opening 6 which creates a connection in the direction of the structured substrate surface 1 with the Containment 4 is connected.
  • This training has particular advantages for the production of the measuring cell 2, since the required recesses are produced with high precision and in one operation can and can be covered with a cover element, which does not have to be structured, without high assembly effort to form the cavities for the containment 4, the measurement opening 6 and the measurement chamber 5.
  • the substance-recognizing component 7 can be any material known from the prior art for carrying out an electrochemical or chemo- / biosensory measurement. All immobilization materials can be used for potentiometric and in particular amperometric biosensors. Examples include gelatin, collagen, alginates, agar, cellulose, triacetate, silicone rubber, polyvinyl alcohol, polyurethane and HEMA. Photocrosslinkable materials can be crosslinked by UV radiation after filling. The active substance-recognizing components such as enzymes or antibodies are immobilized in these materials. This can be done by known methods.
  • the membrane material is contacted by electrically conductive contact surfaces 9, which are attached to the bottom of the recess of the containment 4.
  • the contact surface 9 should be chosen to be as large as possible, particularly in the case of an amperometric measurement principle.
  • the contact surface 9 is connected to the measuring electronics via an electrically conductive contact track 10.
  • the contact surfaces 9 and the contact tracks 10 can be made, for example, by applying thin metallic layers, e.g. by evaporation or
  • Sputtering are applied to the structured substrate surface 1 and / or the cover element.
  • the thickness of these layers is typically approximately 50 to 100 nm. If necessary, corresponding depressions are on to provide the structured substrate surface 1 or the cover element.
  • the electrical contact layers preferably consist of noble metal films such as platinum, gold or silver. However, other electrically conductive materials such as graphite or aluminum can also be used.
  • the electrical contact layers can be applied, for example, by known thin-film technologies, vapor deposition, sputtering, photolithographically structured films, vapor deposition or sputtering with subsequent structuring, vapor deposition or sputtering through shadow masks or using the electrospray method.
  • FIG. 2 shows a section A-A 'of the measuring cell 2 from FIG. 1 with a cover element 11 attached.
  • the cover element 11, like the substrate 3, is preferably plate-shaped. It can be structured on the side opposite the structured substrate surface 1 to form the containment 4, the measuring chamber 5 or the measuring opening 6.
  • the cover element 11 is preferably flat on the side facing the substrate 3.
  • the cover element 11 consists of a suitable material, for example one of the materials proposed above for the substrate 3.
  • the substrate 3 has depressions for the measuring chamber 5, the measuring opening 6 and the containment 4. In the example shown, these depressions are all of the same depth. However, this is not absolutely necessary and can also be designed differently using suitable microstructuring methods, for example in order to achieve an additional cross-sectional constriction in the area of the measuring opening 6 due to a smaller depth.
  • the wells, which . the measuring chamber 5, the containment 4 and the measuring opening form 6 are on the structured substrate surface
  • the membrane material is held securely in the containment 4 and the measurement cell 2 can be produced easily and with high precision.
  • the walls of the depressions are approximately perpendicular to the structured substrate surface 1.
  • the resulting flank angle depends on the manufacturing process used and is preferably between 85 ° and 95 °. However, it can, without affecting the functionality of the measuring cell
  • the cover element 11 is connected to the substrate 3, as a result of which the depressions in the substrate 3 are covered to form cavities.
  • a cover member 11 made of plastic film can be made by a number of known methods such as e.g. thermal lamination, laser welding, gluing, microwave or ultrasonic welding can be connected to the substrate 3.
  • the substance-recognizing constituent 7 can be introduced into the containment 4 before or after the covering element 11 has been applied to the substrate 3. For this purpose, to provide a filling opening in the substrate 3 or the cover element 11.
  • a polymer membrane, a liquid membrane or other relevant membrane materials eg hydrogel
  • containment 4 It is also possible to use a single filling chamber to fill several capillary connecting channels branching off from it to form several containments 4 on a substrate 3. Another possible filling of containment 4 is e.g. the use of an automatic dispensing device based on the inkjet principle.
  • Typical dimensions of a measuring cell according to the invention are as follows: thickness of the substrate approx. 10 ⁇ m to 10 mm, transverse dimensions in the direction of the structured substrate surface 3 approx. 1 mm to 5 cm, depth of the containment 4 approx. 10 ⁇ m to 1 mm and diameter of the containment 4 approx. 10 ⁇ m to 10 mm.
  • FIG. 3 shows a section BB * corresponding to FIG. 2 corresponding to FIG. 1.
  • FIG. 4 shows a measuring cell 2 which differs from the measuring cell shown in FIG. 1 by a different shape of the containment 4.
  • the shape of the containment 4 can be adapted as required to the respective requirements; for example, the recess of the containment 4 can have a circular, triangular or rectangular cross section.
  • the requirements of the membrane material with the substance-recognizing component 7 and the detection technology, the requirements with regard to the measuring chamber 5 and in particular the requirements with regard to a measuring opening 6 forming a constriction can be taken into account.
  • the containment 4 may taper continuously or discontinuously, in the form of a channel, in sections or in any other manner towards the measuring opening 6.
  • FIG. 5 shows a section AA ′ to FIG. 4 with a cover element 11; in this respect it corresponds to FIG. 3.

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Electrochemistry (AREA)
  • Molecular Biology (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

L'invention vise à produire de manière simple et économique une cellule de mesure (2) électrochimique pour effectuer des mesures électrochimiques ou biodétectrices sur des analytes. A cet effet, il est prévu de pratiquer, dans un substrat (5), des évidements pour une chambre de mesure (5), une ouverture de mesure (6) et un espace de confinement (4) contenant le constituant (7) identifiant la substance concernée et de les recouvrir d'un élément de recouvrement (11). Les espaces creux formés par les évidements sont entourés, sur leur face opposée à l'élément de recouvrement (11), par le matériau du substrat (3). Un appauvrissement du constituant (7) identifiant la substance concernée est contrecarré par un rétrécissement de l'ouverture de mesure (6).
PCT/EP2002/004681 2001-05-16 2002-04-27 Cellule electrochimique de mesure de debit WO2002093153A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE10123803.7 2001-05-16
DE2001123803 DE10123803C1 (de) 2001-05-16 2001-05-16 Elektrochemische Messzelle

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Cited By (3)

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WO2010091414A3 (fr) * 2009-02-09 2010-09-30 Forensic Science Service Limited Perfectionnements apportés ou se rapportant à des composants
DE102010064392A1 (de) * 2010-10-29 2012-05-03 Endress + Hauser Conducta Gesellschaft für Mess- und Regeltechnik mbH + Co. KG Verfahren zur Bestimmung eines Analytgehalts einer Flüssigkeitsprobe mittels eines Bioanalysators
DE102014105575A1 (de) * 2014-04-17 2015-10-22 Innovative Sensor Technology Ist Ag Verfahren zur Herstellung einer pH-Halbzelle und eine pH-Halbzelle

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DE102010063031A1 (de) * 2010-12-14 2012-06-14 Endress + Hauser Conducta Gesellschaft für Mess- und Regeltechnik mbH + Co. KG Potentiometrischer Sensor und Verfahren zur Inbetriebnahme eines potentiometrischen Sensors

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Cited By (6)

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Publication number Priority date Publication date Assignee Title
WO2010091414A3 (fr) * 2009-02-09 2010-09-30 Forensic Science Service Limited Perfectionnements apportés ou se rapportant à des composants
US8640555B2 (en) 2009-02-09 2014-02-04 Bioaccel Performance
DE102010064392A1 (de) * 2010-10-29 2012-05-03 Endress + Hauser Conducta Gesellschaft für Mess- und Regeltechnik mbH + Co. KG Verfahren zur Bestimmung eines Analytgehalts einer Flüssigkeitsprobe mittels eines Bioanalysators
DE102010064391A1 (de) * 2010-10-29 2012-05-03 Endress + Hauser Conducta Gesellschaft für Mess- und Regeltechnik mbH + Co. KG Verfahren zur Bestimmung eines Analytgehalts einer Flüssigkeitsprobe mittels eines Bioanalysators
US10036098B2 (en) 2010-10-29 2018-07-31 Endress+Hauser Conducta Gmbh+Co. Kg Method for determining an analyte content of a liquid sample by means of a bioanalyzer
DE102014105575A1 (de) * 2014-04-17 2015-10-22 Innovative Sensor Technology Ist Ag Verfahren zur Herstellung einer pH-Halbzelle und eine pH-Halbzelle

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