JP2014178143A - Concentration measurement sensor, concentration measurement sensor sheet, and manufacturing method thereof - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a concentration measurement sensor, a concentration measurement sensor sheet, and a manufacturing method thereof, capable of preventing a defect caused by presence of endotoxin in an area where a reagent layer is arranged.SOLUTION: A concentration measurement sensor 10 includes: an insulation base material 12 with at least one surface serving as an insulation surface 12a having insulation property; and a conductive pattern 30 arranged on the insulation surface 12a of the insulation base material 12. The insulation base material 12 has a support base material 11 supporting the insulation base material 12 on the other surface thereof. A reagent layer 18 is arranged on the conductive pattern 30. An endotoxin concentration in an area 18a on which at least the reagent layer 18 is arranged, of the insulation base material 12 and the conductive pattern 30, is 0.01EU/mL or lower.

Description

  The present invention relates to a concentration measuring sensor for measuring the concentration of a substance, a sheet for a concentration measuring sensor used for producing such a concentration measuring sensor, and a method for manufacturing the same.

  As a method for quickly and easily measuring a concentration or the like of a specific component in a biological sample such as blood, a sensor (concentration measurement sensor) using an electrochemical detection means has been put into practical use. An example of such a sensor is a glucose sensor that quantifies the glucose concentration in blood electrochemically.

  A glucose sensor includes a base material, an electrode system including a working electrode and a counter electrode provided on the base material, an enzyme, and an electron acceptor as basic components. The enzyme selectively oxidizes glucose in the blood to produce gluconic acid, and at the same time reduces the electron acceptor to produce a reduced form. By applying a certain voltage to the reductant from the external device to the electrode system, the reductant is oxidized again, and current is generated at that time. Since this current value depends on the glucose concentration in the blood, glucose in the blood can be quantified and measured.

JP 2012-127695 A

  By the way, a bacterial wall toxin called endotoxin is generally known. This endotoxin is a toxic substance that constitutes the outer membrane of Gram-negative bacteria such as Escherichia coli and Salmonella. If this endotoxin is mixed in blood or the like even in a very small amount (for example, ng / mL order), it may cause a shock symptom or the like, resulting in the worst death. However, endotoxins are widely present in the air. For this reason, inspections such as the presence of endotoxins in pharmaceuticals such as dialysate are being carried out.

  Conventionally, when measuring the concentration of endotoxin, there are methods called a turbidimetric method and a colorimetric method using a Limulus test. In recent years, methods for measuring the concentration of endotoxin using an electrochemical method have also been studied. For example, a technique is known in which an electrode is placed in a mixture of an analyte and a reagent, and measurement based on differential pulse voltammetry (DPV) is performed (see Patent Document 1).

  Moreover, the concentration measurement sensor which measures glucose concentration and endotoxin concentration using an electrochemical method has a base material and an electrode. In this case, the electrode is generally formed on the substrate by using a screen printing method, a gravure printing method, a vapor deposition method, or the like. However, endotoxins are widely present in the production sites of electrode and substrate materials and concentration measurement sensors. For this reason, in a concentration measurement sensor that measures an endotoxin concentration, endotoxin may exist on the electrode after the electrode is formed on the substrate. Therefore, if the endotoxin on the electrode is not removed, there arises a problem that the endotoxin adhering to the electrode is erroneously measured when measuring the endotoxin. Further, in a concentration measurement sensor that measures glucose concentration, blood is collected by instantaneously attaching the concentration measurement sensor to blood. At this time, endotoxin accidentally attached to the electrode may enter the body and cause problems.

  The present invention has been made in consideration of such points, and a concentration measurement sensor, a concentration measurement sensor sheet, and a concentration measurement sensor capable of preventing problems caused by the presence of endotoxin in the region where the reagent layer is disposed, and It aims at providing the manufacturing method.

  The present invention provides a concentration measurement sensor for measuring the concentration of a substance in a solution, wherein at least one surface is an insulating surface having an insulating property, and the insulating substrate has an insulating surface on the insulating surface. A conductive layer provided and a reagent layer disposed on the conductive pattern, and at least an endotoxin concentration in a region where the reagent layer is disposed is 0 among the insulating base material and the conductive pattern. .01EU / mL or less is a concentration measurement sensor.

  The present invention is the concentration measurement sensor, wherein the insulating substrate has a thickness of 5 μm to 300 μm.

  The present invention is the concentration measurement sensor, wherein the conductive pattern has a thickness of 0.3 μm to 50 μm.

  The present invention is characterized in that at least a region of the conductive pattern in which the reagent layer is arranged includes a base conductive layer and a conductive pattern protective layer provided on the base conductive layer. It is a measurement sensor.

  The present invention is the concentration measurement sensor, wherein a supporting base material that supports the insulating base material is disposed on the other surface of the insulating base material.

  The present invention provides a sheet for a concentration measurement sensor for producing a concentration measurement sensor, wherein at least one surface is an insulating substrate having an insulating property, and is provided on the insulating surface of the insulating substrate. The endotoxin concentration of at least the region where the reagent layer is arranged among the insulating base material and the conductive pattern is 0.01 EU / mL or less. It is the sheet | seat for density | concentration measurement sensors.

  The present invention is the concentration measurement sensor sheet, wherein the insulating base has a thickness of 5 μm to 300 μm.

  The present invention is the concentration measurement sensor sheet, wherein the conductive pattern has a thickness of 0.3 μm to 50 μm.

  The present invention is characterized in that at least a region of the conductive pattern in which the reagent layer is arranged includes a base conductive layer and a conductive pattern protective layer provided on the base conductive layer. It is a sheet for a measurement sensor.

  The present invention is a method for manufacturing a concentration measurement sensor, comprising: an insulating base material supplying step of supplying a strip-shaped insulating base material having at least one surface having an insulating surface; and the insulating base material A conductive pattern forming step of forming a plurality of conductive patterns on the insulating surface, and the endotoxin concentration in the region where at least the reagent layer is arranged among the insulating base material and the conductive pattern is 0.01 EU / It is a manufacturing method of the sheet | seat for density | concentration measurement sensors characterized by setting it as mL or less.

  The present invention further comprises a step of performing a flash annealing process, a plasma ashing process, or a heat sterilization process on the insulating substrate and the conductive pattern. Is the method.

  This invention is a manufacturing method of the sheet | seat for density | concentration measurement sensors characterized by the said insulating base material supply process and the said electroconductive pattern formation process being implemented in a clean room.

  ADVANTAGE OF THE INVENTION According to this invention, the malfunction which arises when endotoxin exists in the area | region where a reagent layer is arrange | positioned can be prevented.

FIG. 1 is an exploded perspective view showing a concentration measuring sensor according to a first embodiment of the present invention. FIG. 2 is a perspective view showing a concentration measurement sensor and an external device. FIG. 3 is an exploded perspective view showing a modified example of the concentration measuring sensor. FIG. 4 is a plan view showing a concentration measuring sensor sheet according to the first embodiment of the present invention. FIGS. 5A and 5B are schematic views showing a concentration measuring sensor manufacturing apparatus. Fig.6 (a) is a top view which shows the insulating base material in an insulating base material supply process, FIG.6 (b) is the VI-VI sectional view taken on the line of Fig.6 (a). FIG. 7A is a plan view showing an insulating substrate in the water-soluble resin layer forming step, and FIG. 7B is a cross-sectional view taken along the line VII-VII in FIG. Fig.8 (a) is a top view which shows the insulating base material in a conductive layer formation process, FIG.8 (b) is the VIII-VIII sectional view taken on the line of Fig.8 (a). Fig.9 (a) is a top view which shows the insulating base material in a water-soluble resin layer removal process, FIG.9 (b) is the IX-IX sectional view taken on the line of Fig.9 (a). FIG. 10A is a plan view showing a support substrate and an insulating substrate in the support substrate disposing step, and FIG. 10B is a cross-sectional view taken along line XX of FIG. Fig.11 (a) is a top view which shows the sheet | seat for density | concentration measurement sensors in a cutting process, FIG.11 (b) is the XI-XI sectional view taken on the line XI of Fig.11 (a). FIG. 12 is a plan view showing a concentration measuring sensor according to a second embodiment of the present invention. FIG. 13 is a plan view showing a sheet for a concentration measurement sensor according to a second embodiment of the present invention.

(First embodiment)
The first embodiment of the present invention will be described below with reference to the drawings. Drawing is an illustration and may exaggerate a characteristic part for explanation, and may differ from an actual thing. In addition, the present invention can be implemented with appropriate modifications without departing from the technical idea. Note that, in the following drawings, the same portions are denoted by the same reference numerals, and some detailed description may be omitted.

(Concentration measurement sensor)
First, an embodiment of a concentration measuring sensor will be described with reference to FIG. FIG. 1 is a diagram showing an embodiment of a concentration measuring sensor according to the present invention. The concentration measurement sensor 10 shown in FIG. 1 measures the concentration of a substance in a solution, and detects a blood glucose level by measuring the concentration of glucose in blood, for example.

  Such a concentration measuring sensor 10 includes a support base 11, an insulating base 12 provided on the support base 11, and a pair provided on the surface (one surface) 12 a of the insulating base 12. Wiring portions 13a and 13b.

  Among these, the insulating base material 12 constitutes a base material whose surface 12a is an insulating surface. Moreover, the insulating base material 12 is hold | maintained by the support base material 11 which has rigidity in the back surface (other side) 12b side.

  A working electrode 14a is provided at one end of one wiring portion 13a, and a counter electrode 14b is provided at one end of the other wiring portion 13b. The working electrode 14a and the counter electrode 14b constitute an electrode system 14 in contact with the solution to be measured.

  Furthermore, connection terminals 15a and 15b are provided at the other ends of the pair of wiring portions 13a and 13b, respectively.

  The connection terminals 15a and 15b are connected to a connection portion (not shown) on the external device 25 side when the concentration measurement sensor 10 is inserted into the insertion port 26 of the external device 25 as will be described later (FIG. 2). ).

  The pair of wiring portions 13a and 13b, the working electrode 14a and the counter electrode 14b, and the pair of connection terminals 15a and 15b constitute a conductive pattern 30.

  A cavity 17 is provided to cover the conductive pattern 30. The cavity 17 is made of an insulating material and has a suction port 17a that guides blood to be measured from the outside to the working electrode 14a and the counter electrode 14b. The cavity 17 covers the conductive pattern 30 so as to expose the working electrode 14a, part of the counter electrode 14b, and part of the pair of connection terminals 15a, 15b to the outside.

  Further, a cover 19 made of an insulating material is provided on the cavity 17, and a reagent layer 18 is provided on the conductive pattern 30 and on the cavity 17. That is, the reagent layer 18 containing glucose oxidase is held by the cavity 17 and the cover 19.

  By the way, in this Embodiment, the endotoxin density | concentration of the area | region 18a where the reagent layer 18 is arrange | positioned among the insulating base material 12 and the electroconductive pattern 30 is 0.01 EU / mL or less. For example, the endotoxin concentration of the electrode system 14 including the working electrode 14a and the counter electrode 14b and the insulating base material 12 positioned around the electrode system 14 is 0.01 EU / mL or less. EU means an endotoxin activity unit, and is an abbreviation of Endotoxin Unit defined by the US Food and Drug Administration (FDA).

  The portion where the endotoxin concentration is 0.01 EU / mL or less is not limited to only the region 18 a where the reagent layer 18 is disposed in the insulating substrate 12 and the conductive pattern 30. For example, it is preferable that the endotoxin concentration around the region 18a where the reagent layer 18 is arranged in the cavity 17 and the cover 19 is also 0.01 EU / mL or less. Further, for example, the endotoxin concentration may be 0.01 EU / mL or less over the entire region of the insulating substrate 12 and the conductive pattern 30 or the entire region of the concentration measurement sensor 10.

  When the endotoxin concentration is 0.01 EU / mL or less, for example, (i) flash annealing treatment, (ii) plasma ashing treatment, or (iii) heat sterilization treatment for the insulating substrate 12 and the conductive pattern 30. May be implemented. Alternatively, (iv) at least a part of the manufacturing process of the concentration measuring sensor 10 may be performed in a clean room of class 10,000 or less. These methods will be described later.

  Various methods can be used for measuring the endotoxin concentration. For example, the concentration measuring sensor 10 is left to stand in an endotoxin-free water for a certain time (for example, 1 day), and then the endotoxin concentration in the water is measured using a known endotoxin measuring device (for example, ET6000 manufactured by Wako Pure Chemical Industries, Ltd.). The endotoxin concentration may be obtained by using the measurement.

  Next, components of each part will be described.

Support base 11
The support base 11 is preferably made of an insulating resin so that it can be applied to a roll-to-roll method. The support base 11 is made of an insulating resin such as polyethylene terephthalate (PET) resin, polyester resin, vinyl chloride resin, polystyrene (PS) resin, polypropylene (PP) resin, and more preferably made of PET. It has become.

  The support base material 11 can reinforce the insulating base material 12 and has only to have a rigidity that does not break due to its own weight during handling. The support base 11 is preferably thicker than the insulating base 12 in order to reinforce the insulating base 12. Specifically, it may be in the range of 100 μm to 1 mm (referred to as 100 μm or more and 1 mm or less; the same applies hereinafter). In addition, the support base material 11 may consist of the structure which bonded the some base material. By providing the support base 11 in this way, the ease of use of the concentration measurement sensor 10 can be increased.

Insulating substrate 12
The insulating substrate 12 is a substrate that supports the conductive pattern 30, and at least the surface 12a on which the conductive pattern 30 is disposed is an insulating surface. The insulating substrate 12 is preferably made of an insulating resin so that it can be applied to a roll-to-roll method. Specifically, for example, a film of polyethylene terephthalate (PET) resin, polyester resin, vinyl chloride resin, polystyrene (PS) resin, polypropylene (PP) resin, or the like can be suitably used.

  The thickness of the insulating substrate 12 is preferably in the range of 5 μm to 300 μm, for example. By setting the thickness of the insulating substrate 12 to 5 μm to 300 μm, the insulating substrate 12 is not removed even when the endotoxin is removed particularly by plasma ashing (described later). Twelve functions can be retained. In addition, it is preferable to match | combine the thermal expansion coefficient of the insulating base material 12 and the support base material 11, and it is good to comprise the insulating base material 12 and the support base material 11 from the same material especially.

  In addition, in the film thickness measurement process mentioned later, in order to make it easy to measure the thickness of the electroconductive pattern 30, it is preferable that the insulating base material 12 consists of transparent resin. In addition, since it is not necessary to use a relatively expensive colored resin as the material of the insulating substrate 12, the manufacturing cost of the concentration measurement sensor 10 can be reduced.

Conductive pattern 30
The conductive pattern 30 is not particularly limited as long as it has conductivity, and is selected from, for example, Ag, Al, Fe, Ni, Cr, Ti, Ta, Pd, or Cu. A metal material may be included. The conductive pattern 30 may have a thickness in the range of 0.3 μm to 50 μm, for example, depending on the forming method. By setting the thickness of the conductive pattern 30 to 0.3 μm to 50 μm, even if the thickness of the conductive pattern 30 is reduced by the plasma ashing process described later, the thickness that functions as the conductive pattern 30 remains. It becomes possible.

Reagent layer 18
The reagent layer 18 includes an enzyme and an electron acceptor. In the present embodiment, the reagent layer 18 includes glucose oxidoreductase as the enzyme, and the concentration of glucose in blood can be measured. However, the reagent layer 18 may contain an enzyme other than glucose oxidase.

  The reagent layer 18 may contain an enzyme other than glucose oxidase, for example, an enzyme that functions as a cholesterol sensor, an alcohol sensor, a scroll sensor, a lactic acid sensor, or a fructose sensor. In this case, as the enzyme used for each sensor, those suitable for the reaction system such as cholesterol esterase, cholesterol oxidase, alcohol oxidase, lactate oxidase, fructose dehydrogenase, xanthine oxidase, amino acid oxidase can be appropriately used.

Cavity 17 and cover 19
The cavity 17 and the cover 19 can be formed from the same material as that of the insulating substrate 12, and the thickness of the cavity 17 and the cover 19 is 100 μm to 300 μm and 50 μm to 100 μm, respectively.

  As shown in FIG. 3 (modification), the conductive pattern 30 may be composed of a plurality of layers. In this case, the conductive pattern 30 includes a base conductive layer 35 located on the insulating substrate 12 side, and a conductive pattern protection provided on the base conductive layer 35 and at least in the region 18a where the reagent layer 18 is disposed. Layer 39 (conductive ashing sacrificial layer). Among these, the base conductive layer 35 is made of, for example, nickel (Ni) and has a thickness of 100 nm to 20 μm. The method for forming the underlying conductive layer 35 is not limited, but for example, vacuum deposition methods such as vapor deposition or sputtering, printing methods such as screen printing or gravure printing, plating such as electrolytic plating or electroless plating The law etc. can be mentioned.

  The conductive pattern protective layer 39 is provided to prevent a problem that the thin base conductive layer 35 is removed together when a plasma ashing process is performed on the concentration measuring sensor 10 as will be described later. It has been. The conductive pattern protective layer 39 has conductivity, and can protect the underlying conductive layer 35 and also ensure a conductive function. Examples of the conductive pattern protective layer 39 include carbon, nickel, palladium, and gold. Examples of the method for providing the conductive pattern protective layer 39 include a plating method, a vapor deposition method, a screen printing method, and an ink jet printing method.

  The conductive pattern protection layer 39 is preferably provided in at least the region of the base conductive layer 35 that is exposed to the outside when performing the plasma ashing process. However, the present invention is not limited to this. For example, the base conductive layer 35 may be provided so as to cover the entire area.

(Concentration measurement sensor sheet)
Next, an embodiment of the density measuring sensor sheet will be described with reference to FIG. FIG. 4 is a view showing an embodiment of a density measuring sensor sheet according to the present invention. The density measuring sensor sheet 50 shown in FIG. 4 is used when the above-described density measuring sensor 10 shown in FIG. 1 is manufactured, and it does not matter whether it is a single wafer or a roll.

  Such a concentration measurement sensor sheet 50 is provided on the support substrate 11, the insulating substrate 12 provided on the support substrate 11, and the surface (one surface) 12 a of the insulating substrate 12. And a plurality of conductive patterns 30.

  In this case, the endotoxin concentration of at least the region 18a in which the reagent layer 18 is arranged in the insulating substrate 12 and the conductive pattern 30 is 0.01 EU / mL or less. For example, the endotoxin concentration of the electrode system 14 including the working electrode 14a and the counter electrode 14b and the insulating base material 12 positioned around the electrode system 14 is 0.01 EU / mL or less. Moreover, it is preferable that the endotoxin density | concentration around the area | region 18a where the reagent layer 18 is arrange | positioned among the cavity 17 and the cover 19 is also 0.01 EU / mL or less. Alternatively, the endotoxin concentration may be 0.01 EU / mL or less over the entire region of the insulating substrate 12 and the conductive pattern 30 or the entire region of the concentration measurement sensor 10.

  Further, a cavity 17 is provided so as to cover each conductive pattern 30. A cover 19 is provided on each cavity 17, and a reagent layer 18 is interposed between each cavity 17 and each cover 19.

  Although not shown in FIG. 4, in the concentration measurement sensor sheet 50 for producing the concentration measurement sensor 10 shown in FIG. 3, at least the region 18 a in which the reagent layer 18 is disposed in the conductive pattern 30 is A base conductive layer 35 and a conductive pattern protective layer 39 provided on the base conductive layer 35 are included.

  In addition, since the structure of each element which comprises the sheet | seat 50 for density | concentration measurement sensors has already been demonstrated, in FIG. 4, the same code | symbol is attached | subjected to the same part as embodiment shown in FIG. 1 and FIG. The detailed explanation is omitted.

(Sheet for concentration measuring sensor and method for manufacturing concentration measuring sensor)
Next, the density measuring sensor sheet and the method for manufacturing the density measuring sensor will be described with reference to FIGS. Specifically, a method for manufacturing the density measurement sensor sheet 50 shown in FIG. 4 and the density measurement sensor 10 shown in FIG. 1 will be described. FIGS. 5A and 5B are schematic views showing a concentration measurement sensor manufacturing apparatus, and FIGS. 6 to 11 show the insulating properties in the respective steps of the concentration measurement sensor sheet and the concentration measurement sensor manufacturing method, respectively. It is the top view and sectional drawing which show the base material 12 or the sheet | seat 50 for density | concentration measurement sensors.

  First, the strip-shaped insulating base material 12 is continuously supplied from the insulating base material supply roll 41 (FIG. 5A) (insulating base material supply step) (FIGS. 6A and 6B).

  Next, the insulating substrate 12 is conveyed to the water-soluble resin layer forming part 42 (FIG. 5A). In the water-soluble resin layer forming portion 42, the water-soluble resin is applied in a pattern on the insulating surface 12a of the insulating base 12 to form the pattern water-soluble resin layer 32. (Water-soluble resin layer forming step) (FIGS. 7A and 7B).

  In this case, the water-soluble resin layer 32 has a shape corresponding to a portion other than each conductive pattern 30. In other words, the water-soluble resin layer 32 is not provided in the portion corresponding to the conductive pattern 30, and the insulating substrate 12 is exposed.

  The water-soluble resin layer 32 is made of, for example, a lift-off material made of a water-soluble vinyl resin, and the thickness thereof can be set to 0.1 μm to 10 μm, for example. Examples of the method for applying the water-soluble resin layer 32 include a printing method such as a screen printing method or a gravure printing method.

  Next, the insulating base material 12 is conveyed to the conductive layer forming part 43 (FIG. 5A). In this conductive layer forming portion 43, a conductive layer 33 made of nickel is formed on the water-soluble resin layer 32 and on the insulating substrate 12 exposed from the water-soluble resin layer 32 (conductive layer forming step) (FIG. 8). (A) (b)).

  The conductive layer 33 is preferably provided over substantially the entire surface of the water-soluble resin layer 32 and the insulating substrate 12 exposed from the water-soluble resin layer 32.

  The conductive layer 33 is made of a conductive material constituting the conductive pattern 30, and may include, for example, a metal material selected from Ag, Al, Fe, Ni, Cr, Ti, Ta, Pd, or Cu. Further, the thickness of the conductive layer 33 can be set to, for example, 0.3 μm to 50 μm. In the case of performing a film thickness measurement process using transmittance described later, the thickness of the conductive layer 33 is set so that the transmittance of light transmitted through the conductive layer 33 and the insulating substrate 12 is 5% or more. It is preferable.

  In addition, as a method of providing the conductive layer 33, for example, a vacuum film forming method such as an evaporation method or a sputtering method, a printing method such as a screen printing method or a gravure printing method, a plating method such as an electrolytic plating method or an electroless plating method, or the like. Can be mentioned. Among these, when the vacuum film forming method is used, the conductive layer 33 can be formed as a thin film, and the material cost can be reduced. In addition, the conductive layer 33 can be formed at a high speed on the long insulating base 12 fed from the insulating base supply roll 41.

  Next, the insulating substrate 12 is transported to the water-soluble resin layer removing unit 44 (FIG. 5A). In the water-soluble resin layer removing unit 44, the water-soluble resin layer 32 is removed with water, and the conductive layer 33 on the water-soluble resin layer 32 is selectively removed. Thereby, the some electroconductive pattern 30 which consists of a part of electroconductive layer 33 is formed on the insulating base material 12 (water-soluble resin layer removal process) (FIG. 9 (a) (b)). In FIG. 9A, six sets of conductive patterns 30 are shown.

  In this case, as shown in FIG. 5A, water is infiltrated into the conductive layer 33 by ejecting shower-like water from the nozzle 44a of the water-soluble resin layer removing portion 44 onto the conductive layer 33, thereby water-soluble. The resin layer 32 may be removed. Alternatively, the water-soluble resin layer 32 may be removed by immersing the water-soluble resin layer 32 together with the insulating base 12 in water.

  In this way, a plurality of conductive patterns 30 are formed on the surface 12a of the insulating substrate 12 by sequentially performing the water-soluble resin layer forming step, the conductive layer forming step, and the water-soluble resin layer removing step (conductive Pattern formation step).

  The conductive pattern forming step is not limited to this. For example, after the insulating base material supplying step, a plurality of conductive patterns 30 are formed on the surface 12a of the insulating base material 12 by a printing method such as gravure printing. May be formed.

  Subsequently, the insulating substrate 12 is conveyed to the film thickness measuring unit 45 (FIG. 5A). The film thickness measuring unit 45 is composed of, for example, a transmission type optical measuring device. In the film thickness measuring unit 45, light is irradiated to at least one conductive pattern 30 formed on the insulating substrate 12, and the thickness of the conductive pattern 30 is measured using the transmitted light. (Film thickness measurement process). Thereby, it can be confirmed whether or not the thickness of the conductive pattern 30 is within a specified range.

  As described above, the thickness of the insulating substrate 12 is in the range of 5 μm to 300 μm, for example. Thus, since the thickness of the insulating base material 12 is reduced, the light transmittance can be increased, and the film thickness measurement unit 45 can accurately measure the film thickness.

  Subsequently, the insulating base material 12 is transported to the support base material disposition portion 46 (FIG. 5A). In this support base material placement portion 46, the support base material 11 that supports the insulating base material 12 is provided on the back surface 12b of the insulating base material 12 (support base material placement step) (FIG. 10A (B)).

  At this time, the support base material 11 is supplied from the support base material supply roll 47 to the support base material disposition unit 46, and the support base material disposition unit 46 is formed of, for example, an adhesive with the support base material 11 being the insulating base material 12. Affixed to the back surface 12b. In addition, as an adhesive agent, a well-known thing can be used.

  Then, the support base material 11 and the insulating base material 12 are wound up by the winding roll 48 (Fig.5 (a)).

  Thereafter, the support base material 11 and the insulating base material 12 are cut into predetermined sizes, whereby the concentration measuring sensor sheet 50 is cut into a single sheet (cutting step) (FIGS. 11A and 11B). ). Each concentration measurement sensor sheet 50 includes one or a plurality (in this case, six sets) of conductive patterns 30.

  In addition, the support base material 11 and the insulating base material 12 supplied from the support base material arrangement | positioning part 46 as it is, without winding up the support base material 11 and the insulating base material 12 on the winding roll 48 (FIG. 4). It may be cut.

  Subsequently, as shown in FIG. 4, the cavity 17 is provided on the conductive pattern 30 of the density measurement sensor sheet 50 (cavity arrangement step).

  Next, the concentration measuring sensor sheet 50 is conveyed to the endotoxin removing unit 61 (FIG. 5B). In the endotoxin removing unit 61, at least the endotoxin in the region 18 a where the reagent layer 18 is disposed is removed from the insulating base 12 and the conductive pattern 30. Specifically, the endotoxin concentration at this location is 0.01 EU / mL or less (endotoxin removal step). In the present embodiment, the entire region of the insulating substrate 12 and the conductive pattern 30 is subjected to endotoxin removal treatment so that the endotoxin concentration is 0.01 EU / mL or less.

  The endotoxin removal part 61 may consist of a plasma ashing processing apparatus, for example. In this case, in the endotoxin removing unit 61 (plasma ashing apparatus), the insulating base 12 and the conductive pattern 30 are subjected to plasma ashing, and the endotoxins on the surfaces of the insulating base 12 and the conductive pattern 30 are removed. For example, by reacting the surfaces of the insulating base material 12 and the conductive pattern 30 with plasma or the like, and decomposing or removing the surfaces of the insulating base material 12 and the conductive pattern 30 in the gas phase, endotoxin is removed together with the surfaces. It may be removed. As shown in FIG. 3, it is preferable to provide a conductive pattern protective layer 39 on the base conductive layer 35 so that the base conductive layer 35 remains when plasma ashing is performed. Thus, by using plasma ashing treatment, endotoxin can be removed in a short time.

  Or the endotoxin removal part 61 may consist of a flash annealing treatment apparatus. In this case, in the endotoxin removing unit 61 (flash annealing apparatus), the insulating base 12 and the conductive pattern 30 are subjected to flash annealing, and endotoxins on the surfaces of the insulating base 12 and the conductive pattern 30 are removed. For example, only the surfaces of the insulating substrate 12 and the conductive pattern 30 are instantaneously heated (for example, 1 μs to 1 ms) to about 600 ° C. to 1200 ° C., and endotoxins on the surfaces of the insulating substrate 12 and the conductive pattern 30 are reduced. It may be sterilized and removed. As described above, by using the flash annealing treatment, it is possible to remove endotoxin in a short time. Thus, the insulating substrate 12 and the conductive pattern 30 are subjected to the flash annealing treatment, so that the endotoxin concentration on the surfaces of the insulating substrate 12 and the conductive pattern 30 is, for example, from 0.5 EU / mL to 10 EU / mL. It can be significantly reduced to 0.01 EU / mL or less.

  Or the endotoxin removal part 61 may consist of a heat sterilization processing apparatus. In this case, in the endotoxin removing unit 61 (heat sterilization processing apparatus), the insulating base 12 and the conductive pattern 30 are heated, and endotoxins on the surfaces of the insulating base 12 and the conductive pattern 30 are sterilized and removed. For example, the surfaces of the insulating substrate 12 and the conductive pattern 30 are heated to about 600 ° C. to 1200 ° C. for a predetermined time (for example, 30 minutes), and endotoxins on the surfaces of the insulating substrate 12 and the conductive pattern 30 are removed. Also good. In addition, a dry method can be mentioned as a heating method. Thus, by using the heat sterilization treatment, it is possible to sterilize without expensive equipment.

  Alternatively, instead of using the endotoxin removing unit 61, the above-described steps (insulating base material supplying step, conductive pattern forming step, film thickness measuring step, and supporting base material disposing step) are performed in a clean room of class 10,000 or less. You may carry out. Further, even when the endotoxin removal process is performed using the above-described endotoxin removal unit 61 (plasma ashing treatment apparatus, flash annealing treatment apparatus, or heat sterilization treatment apparatus), the above steps and the endotoxin removal process are performed in class 10,000 or less. It may be carried out in a clean room.

  Thereafter, as shown in FIG. 4, a reagent layer 18 is provided on the cavity 17 (reagent layer disposing step), and then a cover 19 is provided on the reagent layer 18 so that the reagent layer 18 is placed between the cavity 17 and the cover 19. (Cover disposing step). In this way, the density measuring sensor sheet 50 shown in FIG. 4 is obtained.

  Thereafter, the concentration measuring sensor sheet 10 shown in FIG. 1 is obtained by cutting the sheet 50 for the concentration measuring sensor and separating the sheet 50 for each conductive pattern 30.

  In the above, the endotoxin removing step is provided after the cavity disposing step, but is not limited thereto. For example, it may be provided after the insulating base material supplying step, after the conductive pattern forming step, after the film thickness measuring step, after the supporting base material disposing step, or after the cutting step. Further, the endotoxin removal step may be performed a plurality of times.

  Next, concentration measurement using the concentration measurement sensor 10 will be described.

  First, the concentration measuring sensor 10 is inserted into the insertion port 26 of the external device 25 (FIG. 2). At this time, the connection portion on the external device 25 side comes into contact with the connection terminals 15a and 15b of the concentration measurement sensor 10, respectively. Then, a switch (not shown) in the external device 25 is activated by the concentration measurement sensor 10, and the external device 25 enters a liquid sample suction standby state.

  Thereafter, the user attaches a liquid sample to the suction port 17 a of the cavity 17 of the concentration measurement sensor 10. At this time, the liquid sample is drawn from the suction port 17 a of the cavity 17 by capillary action of the suction port 17 a of the cavity 17. As the liquid sample, for example, a liquid sample derived from a living body such as blood, sweat or urine, a liquid sample derived from the environment, a liquid sample derived from food, or the like is used. For example, when the concentration measurement sensor 10 is used as a blood glucose level sensor, the user punctures his / her finger, palm, arm or the like to squeeze out a small amount of blood, and this blood is used as a liquid sample to suck the suction port of the cavity 17. It adheres to 17a.

  In the present embodiment, as described above, the endotoxin concentration in at least the region 18a in which the reagent layer 18 is disposed is 0.01 EU / mL or less in the insulating substrate 12 and the conductive pattern 30. As a result, when the user instantaneously attaches blood such as his / her finger, palm or arm to the suction port 17a of the cavity 17, the endotoxin attached to the concentration measuring sensor 10 may enter the body. Absent.

  The blood adhering to the suction port 17a of the cavity 17 then reaches the reagent layer 18. After the blood reaches the reagent layer 18, the glucose concentration in the blood is measured by the external device 25, and the measurement result is obtained from the external device 25. It is displayed on the display unit 27.

  Next, the principle of measuring the glucose concentration by the external device 25 will be described.

  First, the reagent layer 18 contains glucose oxidoreductase and potassium ferricyanide as an electron acceptor, specifically reacts with glucose in blood, and emits gluconic acid and electrons. This electron turns potassium ferricyanide into potassium ferrocyanide. By applying a certain voltage from the external device 25 side through the connection terminals 15a and 15b, the wiring portions 13a and 13b, the working electrode 14a and the counter electrode 14b, potassium ferricyanide is added again. Then, current is generated. Since the current value in this case is proportional to the glucose concentration in the blood, the glucose concentration can be measured by measuring this current value with the external device 25.

The reaction in the reagent layer 18 will be further described. When glucose oxidase (GOD) is used as the oxidoreductase and potassium ferricyanide (K ₃ Fe (CN) ₆ ) is used as the electron carrier, the following reaction occurs in the reagent layer 18.

Glucose +2 [Fe (CN) ₆ ] ³⁻ + H ₂ O → Gluconic acid + 2H ⁺ +2 [Fe (CN) ₆ ] ⁴⁻

  At this time, the ferrocyanide ions are oxidized at the working electrode 14a to generate an oxidation current, and are reduced to ferricyanide ions as follows.

^{_{2 [Fe (CN) 6]}} 4- → 2 [Fe (CN) 6] 3- + 2e -

  After the measurement is completed, the user pulls out the concentration measurement sensor 10 from the insertion port 26 of the external device 25.

  As described above, according to the present embodiment, the endotoxin concentration in at least the region 18a in which the reagent layer 18 is disposed is 0.01 EU / mL or less in the insulating base 12 and the conductive pattern 30. As a result, when the user instantaneously attaches blood such as his / her finger, palm or arm to the suction port 17a of the cavity 17, the endotoxin adhering to the concentration measuring sensor 10 enters the body, which causes a problem. There is no risk of causing.

(Second Embodiment)
Next, a second embodiment of the present invention will be described with reference to FIGS. 12 and 13 are views showing a second embodiment of the present invention. 12 and 13, the same parts as those in the first embodiment described above are denoted by the same reference numerals and detailed description thereof is omitted.

  The concentration measuring sensor 10 shown in FIG. 12 is used when measuring the concentration of endotoxin, and the concentration measuring sensor sheet 50 shown in FIG. 13 is used when producing the concentration measuring sensor 10 shown in FIG. It is what

  In the concentration measurement sensor 10 shown in FIG. 12 and the concentration measurement sensor sheet 50 shown in FIG. 13, a reference electrode wiring portion 13c is provided between one wiring portion 13a and the other wiring portion 13b. A reference electrode 14c made of, for example, a silver-silver chloride electrode is provided at one end of the wiring portion 13c, and a connection terminal 15c is provided at the other end of the wiring portion 13c.

  In this case, the reagent layer 18 contains a substance that reacts with endotoxin contained in the solution to be measured. For example, the reagent layer 18 may contain a substance containing factor C, factor B, and a clotting enzyme precursor, specifically, horseshoe crab, amebocyte lysate (a horseshoe crab blood cell extract).

  Also in the present embodiment, the endotoxin concentration of at least the region 18a in which the reagent layer 18 is arranged is 0.01 EU / mL or less in the insulating base material 12 and the conductive pattern 30. Accordingly, it is possible to prevent the endotoxin concentration from being erroneously measured due to the presence of the endotoxin in the region 18a where the reagent layer 18 is disposed in advance.

  12 and 13, the method for setting the endotoxin concentration to 0.01 EU / mL or less is the same as that in the first embodiment described above. That is, for example, (i) flash annealing treatment, (ii) plasma ashing treatment, or (iii) heat sterilization treatment may be performed on the insulating substrate 12 and the conductive pattern 30. Alternatively, (iv) at least a part of the manufacturing process of the concentration measuring sensor 10 may be performed in a clean room of class 10,000 or less.

  The density measurement sensor 10 shown in FIG. 12 and the density measurement sensor sheet 50 shown in FIG. 13 can be manufactured in substantially the same manner as in the first embodiment described above. 12 and 13, the same parts as those in the embodiment shown in FIGS. 1 to 11 are denoted by the same reference numerals, and detailed description thereof is omitted.

  A plurality of constituent elements disclosed in the above-described embodiment can be appropriately combined as necessary. Or you may delete a some component from all the components shown by the said embodiment. For example, the form shown in FIG. 12 and the form shown in FIG. 3 are combined, and also in the concentration measurement sensor 10 shown in FIG. 12, at least the region 18 a in which the reagent layer 18 is disposed in the conductive pattern 30. And a conductive pattern protective layer 39 on the underlying conductive layer 35.

DESCRIPTION OF SYMBOLS 10 Concentration measuring sensor 11 Support base material 12 Insulating base material 13a, 13b Wiring part 14 Electrode system 14a Working electrode 14b Counter electrode 15a, 15b Connection terminal 17 Cavity 18 Reagent layer 19 Cover 30 Conductive pattern 50 Concentration sensor sheet

Claims (12)

In a concentration measurement sensor for measuring the concentration of a substance in a solution,
An insulating base material on which at least one surface is an insulating surface having an insulating property;
A conductive pattern provided on the insulating surface of the insulating substrate;
A reagent layer disposed on the conductive pattern,
An endotoxin concentration in at least a region where the reagent layer is arranged in the insulating base material and the conductive pattern is 0.01 EU / mL or less.   The concentration measuring sensor according to claim 1, wherein the insulating base has a thickness of 5 μm to 300 μm.   The concentration measuring sensor according to claim 1, wherein the conductive pattern has a thickness of 0.3 μm to 50 μm.   The area | region where the said reagent layer is arrange | positioned among the said conductive patterns contains a base conductive layer and the conductive pattern protective layer provided on the said base conductive layer, The Claim 1 thru | or 3 characterized by the above-mentioned. The concentration measuring sensor according to any one of the above.   5. The concentration measuring sensor according to claim 1, wherein a supporting base material that supports the insulating base material is disposed on the other surface of the insulating base material. In the sheet for concentration measurement sensor for producing the concentration measurement sensor,
An insulating base material on which at least one surface is an insulating surface having an insulating property;
A plurality of conductive patterns provided on the insulating surface of the insulating substrate;
An endotoxin concentration in a region where a reagent layer is disposed at least among the insulating substrate and the conductive pattern is 0.01 EU / mL or less, and the concentration measuring sensor sheet.   The thickness of the said insulating base material is 5 micrometers-300 micrometers, The sheet | seat for density | concentration measurement sensors of Claim 6 characterized by the above-mentioned.   The thickness of the said conductive pattern is 0.3 micrometer-50 micrometers, The sheet | seat for density | concentration measurement sensors of Claim 6 or 7 characterized by the above-mentioned.   The region of the conductive pattern in which at least the reagent layer is disposed includes a base conductive layer and a conductive pattern protective layer provided on the base conductive layer. The sheet | seat for density | concentration measurement sensors as described in any one of these. A method for manufacturing a concentration measuring sensor, comprising:
An insulative base material supplying step of supplying a strip-shaped insulative base material in which at least one surface is an insulating surface having an insulating property;
A conductive pattern forming step of forming a plurality of conductive patterns on the insulating surface of the insulating substrate;
A method for producing a sheet for a concentration measurement sensor, wherein an endotoxin concentration in at least a region where a reagent layer is arranged in the insulating substrate and the conductive pattern is 0.01 EU / mL or less.   The concentration measurement sensor sheet according to claim 10, further comprising a step of performing flash annealing treatment, plasma ashing treatment, or heat sterilization treatment on the insulating substrate and the conductive pattern. Production method.   The method for producing a sheet for a concentration measuring sensor according to claim 10 or 11, wherein the insulating base material supplying step and the conductive pattern forming step are performed in a clean room.

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