JP4889015B2 - Sensor electrode body, sensor and sensing system using the same, and method for manufacturing sensor electrode body - Google Patents

Description

  The present invention relates to a sensor electrode body for detecting a target substance, a sensor and a sensing system using the sensor electrode body, and a method for manufacturing the sensor electrode body. The present invention also relates to a sensor electrode body having a thin film layer formed of a dispersion of Prussian blue-type metal complex ultrafine particles, a sensor and sensing system using the same, and a method for manufacturing the sensor electrode body.

  There are various uses of sensors for detecting a target substance, and various modes such as a gas detector for detecting a gas and a sensor for detecting various molecules dissolved in water can be mentioned. Among them, the hydrogen peroxide sensor is important because it is mainly required in the food industry, the medical industry, and the like.

Although hydrogen peroxide is used for disinfection in the food industry, it is preferable that the final product food does not contain hydrogen peroxide. Therefore, a sensor that can detect the presence or absence of hydrogen peroxide with high accuracy is desired.
In the medical industry, a hydrogen peroxide sensor is required as a part of a glucose sensor or the like. When glucose is oxidized by glucose oxidase, hydrogen peroxide is generated as a by-product. By detecting this hydrogen peroxide, the presence or absence of glucose can be detected. Therefore, if the hydrogen peroxide sensing accuracy is improved, the glucose detection accuracy is also improved.

One promising high-precision hydrogen peroxide sensor is an electrochemical sensor using a Prussian blue electrolytic deposition electrode. When the oxidation-reduction reaction is measured with an electrode coated with Prussian blue, the electrochemical characteristics change dramatically depending on the concentration of hydrogen peroxide in the electrolyte. By measuring the current at this time, it can function as a very high performance hydrogen peroxide sensor. Here, the Prussian blue-type metal complex will be described (see FIG. 20). The Prussian blue crystal structure 220 is a basic structure, and in some cases, there are substitutions and defects of transition metals and cyano groups, and various ions and , Water has entered. More specifically, a cyano having a carbon atom 222 and a nitrogen atom 223 between two kinds of metal atoms M ₁ and M ₂ (a metal atom 221 and a metal atom 224 in the figure, respectively) in an NaCl type lattice. It has a three-dimensionally cross-linked structure.

Currently reported sensors using Prussian blue electrodeposited electrodes that have a sensing function with extremely high accuracy have a detection capability of about 0.003 ppm (Non-patent Document 1). On the other hand, currently widely available hydrogen peroxide sensors utilize iodine coulometric titration method, ion transfer spectrometry method, etc., and all have detection performance of about 1 ppm. You can see how high the sensing sensitivity of Prussian blue is.
In addition, the hydrogen peroxide sensor using Prussian blue has a molecular selectivity that does not react with ascorbic acid. When used for medical purposes or food applications, ascorbic acid is often present at the same time, and a sensor capable of detecting only hydrogen peroxide is required, and Prussian blue type complexes are also promising in this respect.

  On the other hand, in recent years, there is a demand for a technique for miniaturizing a sensor and processing it into various shapes. For example, in μ-TAS, a chemical reaction is performed by flowing a material through a thin channel of micrometer or nanometer level to obtain a target product. In order to detect the reactant / product in such a microchannel, a very finely processed sensor is required. In addition, it is required to install a plurality of sensors according to the situation along the flow path, and the need for miniaturization is increasing more and more.

  On the other hand, precise microfabrication is difficult with the electrodeposition electrode as described above. For this reason, it is difficult to apply a sensor using an electrodeposited electrode in a fine space such as a microchannel, and even if fine processing is performed, it takes too much cost and time.

  As an attempt to solve this problem, Non-Patent Document 2 prototypes a molecular sensor using Prussian blue crystals. This sensor has high sensing performance, but requires an extremely complicated process of laminating a polymer layer on the Prussian blue layer. Therefore, it takes 5 days or more for production, and miniaturization is difficult. Moreover, since the particles are only dispersed in water, the range of use and the production methods that can be employed are limited. In addition, a polymer compound may interfere with sensing.

  Prussian blue type complexes are usually weak against alkalis. Therefore, when sensing is performed in a strong alkali, Prussian blue crystals are decomposed, and sensor performance may be lost after several tens of times. When using a sensor for food or medical purposes, the test object may be alkaline, and it is desired to improve the alkali durability of the sensor. If a single-use sensor is used, this problem must be solved in order to make it a sensor of a type that can be used repeatedly. In this regard, attempts have been made to improve the durability of the sensor by preparing a mixture of an insulating polymer and a Prussian blue complex (Non-patent Document 3). However, this method also employs a technique of laminating a polymer layer on the Prussian blue layer, and the manufacturing process is extremely complicated and fine processing is difficult. In addition, the polymer compound may inhibit sensing as described above.

  Recently, methods for obtaining Prussian blue-type metal complexes coated with a low molecular weight compound as fine particles have been studied (see Non-Patent Documents 4 to 8). Here, the synthesis method and basic physical properties of the fine particles are outlined, and the particle size, magnetism, and the like are disclosed, but there is no description of producing a sensor using them.

JP 2004-275168 A JP 2003-274976 A Arkady A. Karyakin et al., Analytical Chemistry, 2004, 76, p474. Pee. A. P. A. Fiorito et al., Chem. Commun., 2005, 366-368. F. F. Ricci et al., Biosensor and Bioelectronics 21 (2005) 389-407 Mami Yamada et al., Journal of the American Chemical Society (J. Am. Chem. Soc), 126 (2004) pp. 9482-9443 Masato Kurihara “Pursuing the Point of Contact between Coordination Chemistry and Its Practical Use”, Hosted by the Chemical Society of Japan, Tohoku Branch (22nd Inorganic and Analytical Chemistry Colloquium) July 1st and 2nd, 2005, Proceedings Mami Yamada "Synthesis and physical properties of Prussian blue Fe / Cr-CN-Co complex nanoparticles using reverse micelle method", sponsored by the Grant-in-Aid for Scientific Research of this school, sponsored by the Chemical Society of Japan (3rd "Molecular Spin" Symposium) January 8-9, 2005, Proceedings, 32, 33 pages N. Bagkar et al., Journal. of. Materials. Chemistry (Journal of Materials Chemistry), 2004, 14, p1430 Toru Kawamoto “Metal Complex Ultrafine Particles: Diverse External Field Induction Phenomena” 5th Molecular Spin Symposium (December 23, 2005)

  An object of the present invention is to provide a sensor electrode body capable of detecting a target substance with high sensitivity and high accuracy, a sensor and a sensing system using the same, and a method of manufacturing the sensor electrode body. Specifically, an excellent electrode body for a sensor that can form a substance-adsorbing thin film layer using ultrafine particles, not just fine particles, and can electrically detect and adsorb a specific substance on the thin film layer, and a sensor using the same Another object of the present invention is to provide a sensing system and a method for manufacturing a sensor electrode body. In addition, a sensor electrode body that can perform precision thinning and ultra-fine processing regardless of a manufacturing method that requires a great amount of time and man-hours, and can detect a trace amount of substance even in a fine and complicated space, It aims at providing the manufacturing method of the sensor and sensing system using, and the electrode body for sensors.

(1) A substance adsorbing thin film layer having a thickness of 1 × 10 ⁻⁹ to 1 × 10 ⁻⁶ m formed of a dispersion containing Prussian blue-type metal complex ultrafine particles having a protective ligand is formed into a conductive structure. A sensor electrode body provided on the body,
The sensor applies a voltage to the conductive structure to electrically control the electrode body, and measures a selective change in electrical characteristics when a target substance is adsorbed on the surface of the substance adsorption thin film layer. The target substance is detected by
A compound containing a pyridyl group or an amino group as the protective ligand is added to the Prussian blue type metal complex crystal having the following metal atom M ₁ and metal atom M ₂ obtained by stirring extraction method. Ultrafine particles having an average particle diameter of 200 nm or less coordinated with one or more kinds,
Further, the protective ligand is removed by performing a heat treatment and / or a cleaning treatment with a cleaning agent at the time of forming the substance adsorbing thin film layer and / or thereafter .
[Metal atom M ₁ : At least one metal atom selected from vanadium, chromium, molybdenum, tungsten, manganese, iron, ruthenium, cobalt, nickel, platinum, rhodium, osmium, iridium, palladium, and copper. ]
[Metal atom M ₂ : at least selected from vanadium, chromium, manganese, iron, ruthenium, cobalt, rhodium, nickel, palladium, platinum, copper, silver, zinc, lanthanum, europium, gadolinium, lutetium, barium, strontium, and calcium One metal atom. ]
(2) The sensor electrode body according to (1), wherein the target substance is a molecule.
(3) The sensor electrode body according to (1) or (2), wherein the target substance is hydrogen peroxide.
(4) The electrode body for a sensor according to any one of (1) to (3), wherein the substance adsorption thin film layer contains an electrochemical property control agent.
(5) The substance adsorbing thin film layer is a multilayer thin film layer having at least a thin film layer formed of the dispersion and a thin film layer containing an electrochemical property control agent (1) to ( The electrode body for sensors according to any one of 4).
(6) The electrode body for a sensor according to any one of (1) to (5), wherein the substance-adsorbing thin film layer does not contain a reverse micelle agent .
(7) The sensor electrode body according to any one of (1) to (6), wherein the compound as the protective ligand has 4 to 100 carbon atoms.
(8) The sensor according to any one of (1) to (7), wherein the compound as the protective ligand is represented by any one of the following general formulas (1) to (3): Electrode body.
(In the formula, R ₁ and R ₂ each independently represent a hydrogen atom or an alkyl group, alkenyl group, or alkynyl group having 8 or more carbon atoms.)
(In the formula, R ₃ represents an alkyl group, alkenyl group, or alkynyl group having 8 or more carbon atoms.)
(In the formula, R ₄ represents an alkyl group, alkenyl group, or alkynyl group having 6 or more carbon atoms, and R ₅ represents an alkyl group, alkenyl group, or alkynyl group.)
(9) The sensor electrode body according to (8), wherein the substituents R _{1 to} R ₄ are alkenyl groups.
(10) The material-adsorbing thin film layer contains the protective ligand compound in a mass ratio in a range of 10 times or less of the Prussian blue-type metal complex, thereby increasing alkali resistance (1) to The electrode body for a sensor according to any one of (9).
(11) The substance adsorbing thin film layer is formed by coating the dispersion by any one of a spin coat film forming method, a Langmuir Blodget film forming method, a spray film forming method, a layer-by-layer film forming method, and a printing method. The electrode body for a sensor according to any one of (1) to (10), which is a liquid film-forming layer having a high surface area adsorption surface.
(12) The sensor electrode body according to any one of (1) to (11), wherein the dispersion liquid is a dispersion liquid of Prussian blue-type metal complex ultrafine particles prepared by a stirring extraction method.
(13) The sensor electrode body according to any one of (1) to (12), wherein the stirring extraction method includes the following steps (A) and (B) .
[Step (A): An aqueous solution containing a metal cyano complex (anion) having the metal atom M ₁ as a central metal and an aqueous solution containing a metal cation of the metal atom M ₂ are mixed, and the metal atom M ₁ and step of precipitating crystals of Prussian blue-type metal complex having a metal atom M _2. ]
[Step (B): A step of mixing the solution obtained by dissolving the protective ligand in a solvent with the Prussian blue-type metal complex crystal prepared in Step (A). ]
(14) A sensor comprising the sensor electrode body according to any one of (1) to (13).
(15) The sensor for food inspection according to (14), wherein a trace amount of hydrogen peroxide in food is detected.
(16) The biosensor according to (14), wherein a biomaterial is detected.
(17) A sensing system comprising a combination of the sensor electrode body according to any one of (1) to (13), a counter electrode, and a reference electrode to detect a target substance in a liquid phase.
(18) In manufacturing the electrode body according to any one of (1) to (13),
More stirring extraction method, a protective ligand comprising a compound containing an amino group or pyridyl group, were dispersed ultrafine particles Prussian blue-type metal complex having a following metal atom M ₁ and metal atom M ₂ Prepare a dispersion,
A material adsorbing thin film layer having a thickness of 1 × 10 ⁻⁹ to 1 × 10 ⁻⁶ m is formed on the conductive structure using the dispersion liquid,
A method for producing an electrode body for a sensor, wherein the protective ligand is removed during and / or after the formation of the substance-adsorbing thin film layer by performing a cleaning treatment with a cleaning agent and / or a heat treatment .
[Metal atom M ₁ : At least one metal atom selected from vanadium, chromium, molybdenum, tungsten, manganese, iron, ruthenium, cobalt, nickel, platinum, rhodium, osmium, iridium, palladium, and copper. ]
[Metal atom M ₂ : at least selected from vanadium, chromium, manganese, iron, ruthenium, cobalt, rhodium, nickel, palladium, platinum, copper, silver, zinc, lanthanum, europium, gadolinium, lutetium, barium, strontium, and calcium One metal atom. ]
(19) S Pinkoto casting method, the Langmuir-Blodgett film-forming method, layer-by-layer dip film forming method, and by coating the dispersion liquid by film forming method selected from printing method to form a material adsorbed film layer (18) The manufacturing method of the electrode body for sensors as described in (18) characterized by the above-mentioned.
(20) The method for producing a sensor electrode body according to (18) or (19), wherein the stirring extraction method includes the following steps (A) and (B) .
[Step (A): An aqueous solution containing a metal cyano complex (anion) having the metal atom M ₁ as a central metal and an aqueous solution containing a metal cation of the metal atom M ₂ are mixed, and the metal atom M ₁ and step of precipitating crystals of Prussian blue-type metal complex having a metal atom M _2. ]
[Step (B): A step of mixing the solution obtained by dissolving the protective ligand in a solvent with the Prussian blue-type metal complex crystal prepared in Step (A). ]

The sensor electrode body of the present invention can detect the adsorption of a substance on the surface of the substance-adsorbing thin film layer with high sensitivity and accuracy by being electrically controlled, and its performance is far superior to the conventional one.
In the sensor electrode body of the present invention, since a thin film is formed with a dispersion containing Prussian blue-type metal complex ultrafine particles having a protective ligand, processing time and man-hours are remarkably reduced, and precision thin film processing can be performed easily and accurately. There is an excellent effect that ultra-fine processing can be performed. Further, in the sensor electrode body of the present invention, since a dispersion liquid in which the ultrafine particles are nano-sized and uniformly and stably dispersed in water or various organic solvents is used, a highly precise and high surface area homogeneous thin film layer can be obtained. Even if it is realized and is a complicated fine space, it can be applied as a high-performance sensor.
Furthermore, in the sensor electrode body of the present invention, the protective ligand coordinated to the ultrafine particles can be appropriately removed at the time of film formation and / or after film formation to adjust the electrochemical responsiveness and the like. Has the advantage. Furthermore, it is possible to achieve both the manufacturing quality during film formation and the product quality related to sensor performance.

The present invention will be described in detail below.
About the electrode body for sensors of this invention, the preferable aspect is typically shown in sectional drawing of FIG. The sensor electrode body shown in FIG. 1 has a substance adsorbing thin film layer 1 provided on one side of a conductive structure 2. In the present invention, the term “conductive structure” means not only a structure in which a conductive film is provided on an insulator such as a substrate, but also includes a conductor or a conductive layer made of only a conductive material without using an insulator. Use. Although the electroconductive material used for the electroconductive structure 2 is not specifically limited, The substance which shows electroconductivity, such as platinum, copper, aluminum, iron, a graphite, indium tin oxide (ITO), can be utilized. The substance adsorbing thin film layer 1 is a thin film layer formed of a dispersion liquid containing nanoparticles (particles on the order of nanometers). Details of the nanoparticles and the substance-adsorbing thin film layer will be described later. In the sensor electrode body of the present invention, in addition to the conductive structure and the substance-adsorbing thin film layer, a functional layer may be provided as necessary. For example, when a biosensor for detecting glucose is used, glucose oxidation A layer such as an enzyme-containing layer may be provided. The sensor electrode body of the present invention can itself be used as a sensor, and if necessary, can be combined with a counter electrode and a reference electrode to form a sensing system. May be.
Since the sensor electrode body of the present invention is excellent in microfabrication, when a microchannel is used after being microfabricated, a conductive layer is attached to the bottom surface or wall surface of the microchannel to form a conductive structure. A substance-adsorbing thin film layer can be provided thereon to provide a road surface coating type sensor. At this time, if the road surface of the microchannel itself has conductivity and functions as a conductive structure, the sensor of the present invention can be obtained simply by providing a coating film layer of an ultrafine particle dispersion having a desired shape on the surface. Can do.
The sensor electrode body of the present invention is not limited to a layered electrode body provided with a substance-adsorbing thin film layer on one side as shown in FIG. 1, and may be one provided with a substance-adsorbing thin film layer on both sides of a flat conductive structure. . In addition, the conductive structure may be a needle-like structure, and may be a needle-like electrode body having a substance-adsorbing thin film layer on the entire outer surface thereof. Also, a substance-adsorbing thin film layer is provided at the tip of the needle-like structure. An electrode body may be used.

The shape of the sensor electrode body of the present invention is not particularly limited as described above, and can be formed by forming into a shape according to the purpose. When a sensor having a desired shape is formed, the sensing region may be formed by using the substance adsorbing thin film layer as a desired shape, and the substance adsorbing thin film layer itself is manufactured in a wide range and a conductive structure (or conductive film) is desired. The sensing area may be limited as the shape.
The size of the sensor electrode body of the present invention is not particularly limited, but when an ultrafine sensor is used, for example, it is preferably 1.0 × 10 ^{−18 to} 1.0 × 10 ⁻¹ m ² . It is preferable to be about 0 × 10 ⁻⁸ m ² . In addition, as will be described later, the substance-adsorbing thin film layer can be a high-surface-area liquid film-forming layer, which enables highly sensitive and stable substance detection regardless of whether it is a large sensor or an ultrafine sensor. Can be. Further, for example, as described above, at least the substance adsorbing thin film layer can be produced in accordance with the μ-Tas fine flow path and the like, and a large number of microfabricated sensors having shapes suitable for various purposes can be provided at necessary places.

The substance that can be detected by the sensor electrode body of the present invention is not particularly limited as long as it is adsorbed on the substance adsorption thin film layer of the sensor electrode body and changes its electrical characteristics. Hydrogen peroxide, NO ₂ ⁻ , S ₂ O ₃ ²⁻ , SO ₃ ²⁻ , hydrazine, dopamine, and the like can be mentioned, and it is preferable to detect molecules. It is more preferable to carry out. Moreover, it is also preferable to indirectly detect and detect a substance involved in the reaction through detection of the detection target substance as in glucose detection described later. The detection method is not particularly limited as long as it is a method for detecting a detection target substance by applying a voltage to a conductive structure and measuring an electrical change under the control thereof, but a method for measuring an electrochemical activity during substance adsorption. It is more preferable to measure the change in potential or current when the target substance is chemisorbed on the substance-adsorbing thin film layer.

The detection and measurement conditions are not particularly limited, and may be a gas phase or a liquid phase, but detection is preferably performed in the liquid phase. The concentration of the detection target substance in the measurement sample is not particularly limited. For example, in the case of hydrogen peroxide, the concentration is preferably 0.002 to 20% by mass. It is preferable that it is 2 * 10 < ^-8 > -20 mass%. According to the sensor electrode body of the present invention, such an extremely small amount of detection is possible.

  According to the sensor electrode body of the present invention, hydrogen peroxide can be detected with high sensitivity as described above, and this can be used as a medical biosensor. As described above, hydrogen peroxide and the like are generated when a biological substance such as glucose is decomposed. Even such a minute by-product can be detected by a sensor using the sensor electrode body of the present invention. For example, it can be used as an excellent biosensor for sensing glucose, cholesterol, lysine and the like. .

The electrode body for sensors of the present invention has a substance adsorption thin film layer formed of a nanoparticle dispersion containing nanoparticles. As the nanoparticles, Prussian blue-type metal complex ultrafine particles having electrochemical response are used.
The Prussian blue-type metal complex ultrafine particles that can be used in the sensor electrode body of the present invention can be schematically illustrated with reference to FIG. That is, in this ultrafine particle 20, the ligand L (22) is coordinated on the surface of the Prussian blue-type metal complex microcrystal 21. However, this figure does not show the relationship in size between the ultrafine crystal and the ligand. Further, the ligand may be upright on the crystal surface as shown in the figure, may be collapsed, or may be in another state.
The coordination amount of the ligand L during the preparation of the ultrafine particles is not particularly limited, and depends on the particle size and shape of the ultrafine particles. For example, the metal atom (metal atom M) in the crystal of the Prussian blue type metal complex The total amount of ₁ and M ₂ is preferably about 5 to 30% in molar ratio (the amount of ligand in the substance-adsorbing thin film layer will be described later). In this way, a stable dispersion liquid (ink for liquid film formation) containing nanoparticles of Prussian blue-type metal complex can be obtained, and a highly accurate substance-adsorbing thin film layer can be produced by liquid film formation. Can do.
As described above, the Prussian blue-type metal complex 21 may have defects and vacancies in its crystal lattice. For example, vacancies are inserted at the positions of iron atoms, and the surrounding cyano groups are substituted with water. It may be. It is also preferable to control the electrochemical characteristics by adjusting the amount and arrangement of the holes.

In the present invention, “ultrafine particles” are nanoparticles (particles micronized in the order of nanometers), and particles that can be dispersed, isolated and redispersed in a variety of solvents in the form of nanoparticles, that is, discrete. Particles (not including those that cannot be isolated from a dispersion or dispersion, or those that cannot be isolated / redispersed). The average particle diameter is preferably 200 nm or less, and more preferably 50 nm or less.
In the present invention, unless otherwise specified, the particle diameter means the diameter of a primary particle not containing a protective ligand, and the equivalent circle diameter (from the image of ultrafine particles obtained by observation with an electron microscope, the projected area of each particle). The value calculated as the diameter of the circle corresponding to Unless otherwise specified, the average particle size refers to the average value obtained by measuring the particle size of at least 30 ultrafine particles as described above. Or you may estimate from the average diameter computed from the half width of the signal from the powder X-ray diffraction (XRD) measurement of the powder of an ultrafine particle.
However, when dispersed in a solvent, a plurality of nanoparticles move as secondary particles in a group, and a larger average particle diameter may be observed depending on the measurement method. When the ultrafine particles are secondary particles in a dispersed state, the average particle size is preferably 200 nm or less. It should be noted that larger aggregated particles may be formed by removal of the protective ligand due to treatment after forming the ultrafine particle film, and the present invention is not construed as being limited thereto.

Examples of the method for producing the Prussian blue type metal complex ultrafine particles include a stirring extraction method, a reverse micelle method, and a method using ferritin as a template . In the present invention, the stirring extraction method is employed. It will be described below in detail 撹拌抽-out method and the reverse micelle method.

<Stirring extraction method>
The stirring extraction method is preferable because it can easily synthesize a large amount of ultrafine particles in which various protective ligands are coordinated on the surface without using a special compound used in the reverse micelle method described later. When showing the specific procedures, for example, a solution of a metal cyanide complex (anion) to a central metal of the metal atom M _1, and a metal cation solution, of which central metal is a metal atom M ₂ were mixed, metal Precipitating a Prussian blue-type metal complex crystal composed of an atom M ₁ and a metal atom M ₂ , then adding the Prussian blue-type complex crystal to a solvent in which the protective ligand L is dissolved, stirring, and removing the solvent. Thus, for example, an aggregate of solid powder ultrafine particles having a particle diameter of 50 nm or less can be obtained.

  The stirring extraction method will be described in more detail. A Prussian blue-type metal complex ultrafine particle dispersion that can be used in the sensor electrode body of the present invention is manufactured in a process including the following (A) and (B), The ultrafine particles are obtained in a process including the following (A) to (C). Further, as a preferred embodiment, when producing the organic solvent-dispersed ultrafine particles, the step (B) is designated as the step (B1), and when producing the water-dispersed ultrafine particles, the step (B) is designated as the step (B2). And Hereinafter, each process will be described in detail.

In the step (A), an aqueous solution containing a metal cyano complex (anion) having the metal atom M ₁ as a central metal and an aqueous solution containing a metal cation of the metal atom M ₂ are mixed, and the metal atom M ₁ and a step of precipitating crystals of Prussian blue-type metal complex having a metal atom M _2.

The metal atom _{M 1} is vanadium (V), chromium (Cr), molybdenum (Mo), tungsten (W), manganese (Mn), iron (Fe), ruthenium (Ru), cobalt (Co), nickel (Ni ), Platinum (Pt), copper (Cu), rhodium (Rh), osmium (Os), iridium (Ir), palladium (Pd) and the like, and at least one of them is preferable.

As the metal atom _{M 2} is vanadium (V), chromium (Cr), manganese (Mn), iron (Fe), ruthenium (Ru), cobalt (Co), rhodium (Rh), nickel (Ni), palladium (Pd) , Platinum (Pt), copper (Cu), silver (Ag), zinc (Zn), lanthanum (La), europium (Eu), gadolinium (Gd), lutetium (Lu), barium (Ba), strontium (Sr) , Calcium (Ca) and the like, and at least one of them is preferable.

Of these iron as the metal atom M _1, chromium or cobalt is preferable, iron is particularly preferred. Iron as the metal atom M ₂ is comprised of cobalt, nickel, vanadium, copper, manganese or preferably zinc, iron, cobalt or nickel, is more preferable.

But not limited counterion especially a metal atom M ₁ in anionic metal-cyano aqueous solution of a complex of a central metal, potassium ions, ammonium ions, sodium ions, and the like. Although counter ion in an aqueous solution of metal cations the metal atom _{M 2} is not particularly ^{_{limited, Cl -, NO 3 -,}} SO 4 2- , and the like.

Two or more metals may be combined as the metal atom M ₁ or M ₂ . Regarding the combination of two kinds of metals, the metal atom M ₁ is preferably a combination of iron and chromium, a combination of iron and cobalt, a combination of chromium and cobalt, and more preferably a combination of iron and chromium. For the metal atom M _2, a combination of iron and nickel, a combination of iron and cobalt, preferably a combination of nickel and cobalt, a combination of iron and nickel is more preferable. At this time, it is preferable to control the physical properties of the obtained Prussian blue-type metal complex ultrafine particles by adjusting the composition of the combined metals, and it is more preferable to control the electrochemical characteristics thereof.

At this time, the mixing ratio of the metal cyano complex and the metal cation is not particularly limited, but the mixing is preferably performed so that the molar ratio of “M ₁ : M ₂ ” is 1: 1 to 1: 1.5.

Step (B) is a step of mixing a solution in which the protective ligand L is dissolved in a solvent and the Prussian blue-type metal complex crystal prepared in step (A).
As the protective ligand, it is preferable to use one or two or more compounds having a pyridyl group or an amino group as a bonding site with the crystal particle. Among them, a compound having 4 to 100 carbon atoms (mass average molecular weight) It is more preferable to use 2000 or less compounds, and 50 to 1000 compounds are more preferable. One or two of the compounds represented by any one of the following general formulas (1) to (3) It is particularly preferred to use more than one species.

In General Formula (1), R ₁ and R ₂ each independently represent a hydrogen atom or an alkyl group, an alkenyl group, or an alkynyl group having 8 or more carbon atoms (preferably 12 to 18 carbon atoms). R ₁ and R ₂ are preferably alkenyl groups, and the number of carbon-carbon double bonds is not particularly limited, but is preferably 2 or less. When the ligand L having an alkenyl group is used, the dispersibility is improved even when it is difficult to disperse in a solvent other than a polar solvent (methanol or acetone that may be eliminated from the ligand, such as chloroform). be able to. Specifically, by using a ligand having an alkenyl group, it can be well dispersed in a nonpolar solvent (for example, hexane) unless the ligand is eliminated. The same applies to R ₃ and R ₄ .
Among the compounds represented by the general formula (1), 4-di-octadecylaminopyridine, 4-octadecylaminopyridine and the like are preferable.

In the general formula (2), R ₃ represents an alkyl group, an alkenyl group, or an alkynyl group having 8 or more carbon atoms (preferably 12 to 18 carbon atoms). R ₃ is preferably an alkenyl group, and the number of carbon-carbon double bonds is not particularly limited, but is preferably 2 or less. Among the compounds represented by the general formula (2), oleylamine is preferable as a ligand having an alkenyl group, and stearylamine is preferable as a ligand having an alkyl group.

In the general formula (3), R ₄ is an alkyl group, alkenyl group, or alkynyl group having 6 or more carbon atoms (preferably 12 to 18 carbon atoms), and R ₅ is preferably (preferably 1 to 60 carbon atoms). A) an alkyl group, an alkenyl group, or an alkynyl group. R ₄ is preferably an alkenyl group, and the number of carbon-carbon double bonds is not particularly limited, but is preferably 2 or less.
In addition, the compound represented by any one of the general formulas (1) to (3) may have a substituent as long as the effect of the present invention is not hindered.

  At this time, the solvent for dissolving the protective ligand L is preferably determined by a combination with the ligand L or the like, and more preferably a solvent that sufficiently dissolves the ligand L in the solvent. When an organic solvent is used as the solvent, toluene, dichloromethane, chloroform, hexane, ether, butyl acetate and the like are preferable. When using a ligand that dissolves in water, such as 2-aminoethanol, water can be used as a solvent, and water-dispersible Prussian blue-type metal complex ultrafine particles can also be obtained. At this time, it is also preferable to use alcohol as a solvent.

  Although the amount of the solvent used at this time is not particularly limited, for example, it is preferable that “ligand L: solvent” is 1: 5 to 1:50 by mass ratio. Moreover, it is preferable to stir when mixing, whereby a dispersion liquid in which the ultrafine particles of Prussian blue-type metal complex are sufficiently dispersed in an organic solvent is obtained.

The addition amount of the ligand L is expressed by a molar ratio of “(M) with respect to metal ions (total amount of metal atoms M ₁ and M ₂ ) contained in the microcrystal of the Prussian blue-type metal complex prepared in the step (A). ₁ + _M 2): L "is 1: 0.2 to 1: is preferably 2.

  In step (B1), an organic solvent-dispersed ultrafine particle is mixed with an organic solution in which ligand L is dissolved in an organic solvent and the Prussian blue-type metal complex crystal prepared in step (A). It is a process to do. In addition, it is preferable to add water in this process at the point which can raise the production | generation speed | rate of a Prussian blue type metal complex ultrafine particle, The addition amount is "solvent: water" 1: 0.01-1 by mass ratio. : It is preferable that it is 0.1.

In the step (B2), when preparing the water-dispersed ultrafine particles, the water-soluble ligand L is alcohol (the alcohol includes lower alcohols such as methanol, ethanol, propanol, etc., and methanol is preferred). And / or a step of mixing a solution dissolved in a solvent composed of water and the Prussian blue-type metal complex crystal prepared in step (A).
Here, when water is added to the solid material obtained after the alcohol separation, an aqueous dispersion in which ultrafine particles of Prussian blue-type metal complex are dispersed in water can be obtained. Further, the Prussian blue-type metal complex crystal prepared in the step (A) is directly added to the aqueous solution of the ligand L and stirred to obtain a dispersion in which the ultrafine particles of the Prussian blue-type metal complex are dispersed in water. Obtainable. However, it is preferable to use an alcohol solvent from the viewpoint of stability and yield of the obtained Prussian blue-type metal complex ultrafine particles.

  Step (C) is a step of separating Prussian blue from the solvent. For example, when the ultrafine particles of Prussian blue type metal complex are dispersed in the solvent, the solvent can be distilled off under reduced pressure to separate the dispersion. The solvent may be removed by filtration or centrifugation as an untreated state. At this time, when the mixed solution is obtained through the step (B1), the solid powder of the ultrafine particle aggregate is obtained by removing the organic solvent. When a mixed solution is obtained through the step (B2), a solvent composed of alcohol and / or water is removed and separated to obtain a water-dispersible ultrafine particle aggregate as a solid powder.

In addition, when manufacturing Prussian blue-type metal complex ultrafine particles that can be used in the sensor electrode body of the present invention, additives may be added as appropriate, and different physical properties from Prussian blue-type metal complex ultrafine particles due to the action of the additives. Can also be given. For example, it is preferable to add ammonia, pyridine, a combination thereof, or the like as a crystal property modifier during the production of Prussian blue-type metal complex ultrafine particles, and control the characteristics of the product depending on the presence or absence and amount of the addition. At this time, it is preferable to add a crystal property modifier in the step (A). Amount of crystal physical property modifier is not particularly limited, the metal atom M _2, is preferably added so as to be 10 to 200 percent by mole ratio.

Furthermore, as described above, a plurality of kinds of metals can be used in combination as the metal atom M ₁ or M ₂ , but the physical properties of Prussian blue type metal complex ultrafine particles can be adjusted by adjusting the composition of the metal. Can be controlled. Specifically, for example, (Fe _1-x Ni _x ) ₃ [Fe (CN) ₆ ] ₂ is used in combination with metal atom M ₂ and iron and nickel, and the composition (“x” in the formula) is adjusted. It is preferable to control.
At this time, Prussian blue-type metal complex ultrafine particles having the target metal composition can be obtained by adding the raw material compound containing the target metal in the step (A) while changing the mixing ratio. In addition, about the stirring extraction method, Japanese Patent Application No. 2006-030481, international patent application PCT / JP2006-302135 specification, etc. can also be referred. The Prussian blue-type metal complex obtained by the stirring extraction method shows excellent dispersion stability in various solvents (in the present invention, “solvent” is used to mean “dispersion medium”) (for example, several A stable dispersion state can be maintained even after a month or more.), An excellent sensor suitable for thinning can be produced.

<Reverse micelle method>
The reverse micelle method is a method for preparing ultrafine complex particles covered with an alkyl protecting agent, and includes the following three steps.
A first reverse micellar solution containing a metal cyano complex (anion) which: (i) a metal atom M ₁ as a central metal, and a second reverse micellar solution containing a metal錯陽ion as a central metal of the metal atom M ₂ The process of preparing each.
(Ii) A step of mixing the first reverse micelle solution and the second reverse micelle solution.
(Iii) A step of adding the long-chain alkyl protecting agent L to the mixed solution obtained in the step (ii) and isolating the obtained nanocrystals.

The metal atom M ₁ , metal atom M ₂ , counter ion, and protective ligand L that can be used in the reverse micelle method are the same as those described in the stirring extraction method, and the preferred ones are also the same. Hereinafter, each of the above steps will be described in more detail.

Step (i)
In this step, a first reverse micelle solution containing a metal complex anion having the metal atom M ₁ as a central metal and a second reverse micelle solution containing a metal atom M ₂ cation are prepared.

It is preferable to prepare a first solution containing a metal complex anion, of which central metal is pre-M ₁ as a first reverse micellar solution, the first solution is obtained by dissolving the above metal complex anion in water . The concentration of the first solution (the number of moles of metal atoms relative to the total amount of the solution) is preferably 0.1 to 1 mol / L. Next, a first reverse micelle solution is obtained by mixing the first solution and an organic solvent (cyclohexane, hexane, isooctane, etc.) in which the reverse micelle agent is dissolved. Examples of the reverse micelle agent include AOT (di-2-ethylhexylsulfosuccinate sodium salt) or NP-5 (polyethylene glycol mono-4-nonylphenyl ether). The amount of the reverse micelle agent used may be a concentration so that the first solution is solubilized as reverse micelles, but in general, the molar ratio of water to the reverse micelle agent (w = [Water] / [AOT or NP-5]) is preferably 5-50.

In the case of the second reverse micelle solution, a second solution containing a cation of the metal atom M ₂ is prepared in the same manner as in the first solution, and this is mixed with an organic solvent in which the reverse micelle agent is dissolved. A second reverse micelle solution can be obtained. The second solution is preferably an aqueous solution of a metal salt (CoCl ₂ , Fe (NO ₃ ) _3, etc.) containing a metal atom M ₂ . The concentration of the second solution (number of moles of metal atoms relative to the total amount of the solution), the type and amount of reverse micelle agent used, and the preferred range of the organic solvent are the same as in the case of the first reverse micelle solution. The volume of the organic solvent is not particularly limited, but is preferably about 10 to 100 ml.

Step (ii)
In this step, the first reverse micelle solution and the second reverse micelle solution are mixed. By this mixing, metal complex nanocrystals are formed. The production rate of nanocrystals can be adjusted by the concentration of the above solution, the concentration of the reverse micelle agent, and the like. The mixing method is not particularly limited, and a normal mixing apparatus can be used. It is preferable that the mixing ratio of both is such that the metal complex anion: metal cation = 1: 0.7 to 1.3 in molar ratio.

Step (iii)
In this step, the protective ligand compound is added to the mixed solution obtained in step (ii). The protective ligand is as described in detail in the stirring extraction method.

  By these ultrafine particle synthesis methods, ultrafine particles of a nanometer-scale metal complex protected by a predetermined protective ligand can be produced.

Furthermore, the substance adsorption thin film layer of the electrode body for sensors of the present invention will be described in detail.
In the sensor electrode of the present invention, the adsorption of thin layers is a thin film layer formed by nanoparticles dispersion containing ultrafine particles as described above, were subjected to heat treatment, washing treatment or the like thereto. It is not necessary that the individual ultrafine particles maintain their shape in the thin film, and the ligand L may be removed. In addition, a different kind of material may be included for improving the electrochemical characteristics. For example, even if an electrochemical characteristic control agent such as ferrocene or Nafion is included, the layer and nanoparticle containing the electrochemical characteristic control agent are included. It is good also as a multilayer thin film layer which consists at least of the layer in which it was contained. In addition, it is preferable that a high molecular compound is not included in a substance adsorption thin film layer.

The method for forming the substance-adsorbing thin film layer is not particularly limited, but it is preferably a liquid film forming thin film layer formed by coating the dispersion liquid (ink) containing the Prussian blue type metal complex ultrafine particles. Specifically, the coating film forming method includes spin coating film forming method, Langmuir Blodget film forming method, spray film forming method, layer-by-layer (LbL) dipping method, and printing method (inkjet printing method, screen printing method, transfer method). Method, letterpress printing method, soft lithography printing method, etc.). This makes it possible to produce a highly accurate microfabricated film. For example, when the film thickness is controlled by the size of a single fine particle during film formation, it is preferable to use the Langmuir Blodget film formation method. When a plurality of types of sensors are spatially installed, it is preferable to use a printing film forming method using a plurality of inks such as inkjet printing.
In addition, it is also preferable to use a laminated chemical bond film forming method. Here, the laminated chemical bond film-forming method is to form ultrafine particles in one layer or several layers, and when this is repeated, the substrate and ultrafine particles or the ultrafine particles are bonded by chemical bonds such as coordinate bonds. It is a film-forming method that bonds and strengthens the film structure.

The thickness of the substance-adsorbing thin film layer is not particularly limited, but is preferably 1 × 10 ⁻⁹ to 1 × 10 ⁻⁶ m, and more preferably 3 × 10 ^{−9 to} 5 × 10 ⁻⁷ m. Further, the substance adsorption thin film layer is preferably a high surface area film suitable for adsorption of a target substance. In this way, stable detection with ultra-high sensitivity becomes possible. At this time, it is possible to increase the surface area of the substance adsorption surface by using a conductive structure having an uneven surface.

In the sensor electrode of the present invention, when forming the material adsorbed film layer by dispersion which contains the Prussian blue-type metal complex ultrafine and / or after cleaning with treatment agents such as acetone For example, heating processing (preferably heat treatment 100 to 150 ° C.) to facilities. By such treatment, the protective ligand can be removed, and for example, the electrochemical response in substance detection can be improved. Therefore, when forming a film, a very large amount of protective ligand is used to stably disperse the ultrafine particles in the solvent to ensure good film-forming properties. After film formation, the sensor performance is improved (for example, response speed and repetition) The ligand L can be removed and adjusted to improve resistance and the like. Thereby, improvement of manufacturing quality and improvement of product quality can be realized at the same time.

In the sensor electrode body of the present invention, even when an acid or an alkaline substance such as acetic acid is contained in the sample to be detected, it is possible to sense the target substance while maintaining its durability. is there. The reason why Prussian blue has particularly low resistance to alkali is considered to be because hydroxyl groups (OH ⁻ ) penetrate into the material, react with Fe to produce Fe (OH) _{3 and} are eluted from the material. Therefore, it is possible to improve alkali resistance by reducing penetration of hydroxyl groups. That is, in the sensor electrode body of the present invention, the Prussian blue-type metal complex can be covered with a desired protective ligand (coating molecule). Can be improved. For example, when a compound having a negatively charged site is used as the coating molecule, the penetration of a hydroxyl group that is an anion can be inhibited.

When the Prussian blue-type metal complex ultrafine particles are contained in the substance adsorbing thin film layer, the protective ligand L contained in the same layer is not coordinated to the Prussian blue-type metal complex, but the coordination bond is lost. It may be a free molecule. The amount of the protective ligand L contained in the substance-adsorbing thin film layer is not particularly limited, and may be adjusted and removed by the above-described heating / washing process during and / or after film formation. When the amount is too large, for example, the content of the protective ligand is preferably 10 times (molar ratio) or less with respect to the Prussian blue-type metal complex. When the amount is reduced to control the device performance, the content is 1 time. The ratio (molar ratio) is more preferably 1/10 or less (molar ratio).
When a desired protective ligand is contained in order to improve the alkali resistance described above, the content of the protective ligand in the substance-adsorbing thin film layer may be appropriately adjusted depending on the type of the protective ligand, the alkali compound, etc. However, it is preferable to contain, for example, 0.1 to 10 times the protective ligand compound part of the Prussian blue-type metal complex ultrafine particles by mass ratio.
There is no particular lower limit to the content of the protective ligand L, but it may inevitably remain (for example, about 1/100 in terms of the molar ratio) when it is removed by, for example, the heating / washing treatment.

  In the sensor electrode body of the present invention, a film forming method that is inexpensive and suitable for ultra-fine processing can be used, such as spin coating, thereby improving the manufacturing cost of the sensor, improving the quality, and improving the detection sensitivity and accuracy. Can be expected. Furthermore, since the sensor can be manufactured by a simple process, it is suitable for mass production of sensors. In addition, since ultrafine particles are used as the sensing material, a flexible sensor that can be bent or a sensor having a complicated shape can be obtained.

  EXAMPLES Hereinafter, although this invention is demonstrated in more detail based on an Example, this invention is limited to these and is not interpreted.

(Preparation Example 1)
An aqueous solution synthesis of (A) Prussian blue bulk material of _{(NH 4) 4 [Fe (} CN) 6] 1.0g were dissolved in _water, Fe a _{(NO 3) 3 · 9H 2} O 1.4g were dissolved in water The aqueous solution was mixed to precipitate Prussian blue microcrystals. Centrifugation separated Prussian blue microcrystals that were insoluble in water, which were washed three times with water and then twice with methanol and dried under reduced pressure.

The result of analyzing the produced Prussian blue bulk with a powder X-ray diffractometer is shown in a chart 31 of FIG. This coincided with the Prussian blue peak retrieved from the standard sample database (see the peak diagram shown in FIG. 3 (I)). In the FT-IR measurement, a peak due to Fe—CN stretching vibration appears in the vicinity of 2070 cm ⁻¹ (see spectrum 41 in FIG. 4), indicating that this solid is Prussian blue.

(B1) Preparation of organic solvent-dispersed Prussian blue ultrafine particles As ligand L, 0.5 ml of water was added to 5 ml of a toluene solution in which a ligand oleylamine containing a long-chain alkyl group was dissolved. 0.2 g of Prussian blue bulk material synthesized in (A) was added to the above solution. When stirred for one day, a deep blue dispersion in which all Prussian blue microcrystals were dispersed in the toluene phase was obtained. When the water and toluene phases were separated and the dark blue toluene phase was filtered, a dispersion of Prussian blue fine particles was obtained (the results of FT-IR measurement at this time are shown in chart 43 in FIG. 4). The measurement result of the ultraviolet-visible absorption spectrum of this dispersion is shown in the spectrum 51 of FIG. The peak around 680 nm is known as Prussian blue Fe-Fe charge transfer absorption, and it can be seen that this fine particle dispersion contains Prussian blue. Moreover, the result of having observed the dispersion liquid of the Prussian blue ultrafine particle which has an oleylamine as this protective ligand with the transmission electron microscope is shown in FIG. From this, it was confirmed that ultrafine particles having an average particle diameter of about 10 to 15 nm were synthesized. At this time, the Prussian blue bulk was not left in the water, but was almost completely extracted as ultrafine particles in the toluene phase.

(C1) Separation and redispersion of solid powder of Prussian blue ultrafine particle aggregate Toluene in the dispersion obtained in (B1) was dried under reduced pressure to obtain a solid powder almost quantitatively. The obtained solid powder was easily redispersed in an organic solvent such as dichloromethane, chloroform, or toluene, and became a deep blue transparent dispersion.
As a result of analyzing the obtained Prussian blue ultrafine particle solid powder with a powder X-ray diffractometer, it coincided with the peak position of Prussian blue contained in the standard sample database (see chart 33 in FIG. 3). The low-angle peak is a broad background due to excessive oleylamine.

(Preparation Example 2)
(A) Synthesis of Prussian blue bulk body In the same manner as in Preparation Example 1, a Prussian blue bulk body was synthesized.
(B2) Preparation of water-dispersed Prussian blue ultrafine particles 0.2 g of Prussian blue bulk material synthesized in (A) was added to 5 ml of methanol solution in which 2-aminoethanol was dissolved as ligand L for about 3 hours. By stirring, a dispersion of Prussian blue-type metal complex ultrafine particles was prepared. The ultrafine particles existed as a solid matter without being dissolved in methanol even after stirring. The result of the FT-IR measurement at this time is shown in the chart 42 of FIG.

(C2) Separation and re-dispersion of solid powder of Prussian blue ultrafine particle aggregate The solid powder α was obtained by removing methanol from the dispersion. When water was added to the solid powder α, everything was dispersed and became a deep blue transparent dispersion β.

  FIG. 7 shows the results of observation of the obtained dispersion β of Prussian blue ultrafine particles having 2-aminoethanol as a protective ligand (the solvent is water) with a transmission electron microscope. From this, it was confirmed that ultrafine particles having an average particle diameter of about 10 to 15 nm were synthesized. Also in the ultraviolet-visible absorption spectrum measurement of this dispersion, a peak showing Prussian blue at 680 nm was observed as in the case of the organic solvent ultrafine particle dispersion (see spectrum 52 in FIG. 5). From these measurement results, it can be seen that an aqueous dispersion of Prussian blue ultrafine particles was obtained.

  The Prussian blue peak was confirmed from the result of analyzing the solid powder α with a powder X-ray diffractometer (see spectrum 32 in FIG. 3). As a result of the peak half-width analysis, the average particle size of the crystals was about 10 nm to 20 nm. This shows that the solid powder α is an aggregate of Prussian blue nanoparticles.

(Preparation Example 3)
To 1.5 ml of an aqueous solution of potassium ferricyanate, K ₃ [Fe (CN) ₆ ] 0.329 g (0.999 mmol), 0.1 ml of aqueous ammonia NH ₃ (28.0%, 14.8N) was added, and 1.0 ml of an aqueous solution of 0.437 g (1.50 mmol) of cobalt nitrate Co (NO ₃ ) ₂ .6H ₂ O was added and stirred for about 3 minutes. Thereafter, Prussian blue-type metal complex crystals were obtained as a red precipitate by centrifugation. This crystal precipitate was washed three times with water and once with methanol. The yield was 0.631 g, and the yield was 105% (over 100% is considered to be an error due to insufficient drying and containing water).
The Prussian blue-type metal complex crystal (cobalt iron cyano complex crystal) obtained in the previous synthesis was added to 3.0 ml of a toluene solution of 0.443 g (1.66 mmol, 100% of the total metal amount (molar ratio)) of oleylamine. Aggregate 0.204g (0.340mmol) was added and stirred for about 1 day. Thus, Prussian blue type metal complex ultrafine particles were obtained in the dispersion. A dispersion sample γ was obtained under the condition of adding ammonia.

Next, the Prussian blue-type metal complex ultrafine particles were separated and obtained as an agglomerated solid by removing the toluene in the dispersion sample γ by drying under reduced pressure.
The dispersion sample γ was centrifuged, a part of the supernatant was taken out, diluted with toluene, and UV-vis optical measurement was performed. The absorption maximum in the visible region of this sample is present at 480 nm (see FIG. 8), which is different from the maximum position of 520 nm obtained without adding ammonia. This result shows that Prussian blue-type metal complex ultrafine particles having different physical properties can be obtained by adding ammonia.

(Preparation Example 4)
(Synthesis of (Fe _0.2 Ni _0.8 ) ₃ [Fe (CN) ₆ ] ₂ )
An aqueous solution (2 ml) of K ₃ [Fe (CN) ₆ ] (0.658 g (2.00 × 10 ⁻³ mol)) was added to FeSO ₄ .7H ₂ O (0.167 g (6.00 × 10 ⁻⁴ mol)). ) aqueous solution) and _{(0.4ml) Ni (NO 3)} 2 · 6H 2 O ( added with stirring in a mixed solution of an aqueous solution of ^{0.698g (2.40 × 10 -3 mol)} ) (1.6ml) It was. The precipitate was centrifugally washed three times with distilled water, and once centrifugally washed with methanol, and then air-dried to obtain Prussian blue-type metal complex crystals having a target metal composition.

(Synthesis of Fe _0.4 Ni _0.6 ) ₃ [Fe (CN) ₆ ] ₂ )
An aqueous solution (2 ml) of K ₃ [Fe (CN) ₆ ] (0.658 g (2.00 × 10 ⁻³ mol)) was added to FeSO ₄ .7H ₂ O (0.334 g (1.20 × 10 ⁻³ mol). )) And an aqueous solution (1.2 ml) of Ni (NO ₃ ) ₂ .6H ₂ O (0.523 g (1.80 × 10 ⁻³ mol)) added with stirring. It was. The precipitate was centrifugally washed three times with distilled water, and once centrifugally washed with methanol and then air-dried to obtain Prussian blue-type metal complex crystals having the target metal composition.

(Synthesis of (Fe _0.6 Ni _0.4 ) ₃ [Fe (CN) ₆ ] ₂ )
An aqueous solution (2 ml) of K ₃ [Fe (CN) ₆ ] (0.658 g (2.00 × 10 ⁻³ mol)) was added to FeSO ₄ .7H ₂ O (0.500 g (1.80 × 10 ⁻³ mol)). )) And an aqueous solution (0.8 ml) of Ni (NO ₃ ) ₂ .6H ₂ O (0.349 g (1.20 × 10 ⁻³ mol)) added with stirring. It was. The precipitate was centrifugally washed three times with distilled water, and once centrifugally washed with methanol and then air-dried to obtain Prussian blue-type metal complex crystals having the target metal composition.

(Synthesis of (Fe _0.8 Ni _0.2 ) ₃ [Fe (CN) ₆ ] ₂ )
An aqueous solution (2 ml) of K ₃ [Fe (CN) ₆ ] (0.658 g (2.00 × 10 ⁻³ mol)) was added to FeSO ₄ .7H ₂ O (0.667 g (2.40 × 10 ⁻³ mol)). ) aqueous solution) and _{(1.6ml) Ni (NO 3)} 2 · 6H 2 O ( added with stirring to a mixed solution of aqueous solution (0.4 ml) of ^{0.174g (5.98 × 10 -4 mol)} ) It was. The precipitate was centrifugally washed three times with distilled water, and once centrifugally washed with methanol and then air-dried to obtain Prussian blue-type metal complex crystals having the target metal composition.

To 0.10 g of the four kinds of (Fe _1-x Ni _x ) ₃ [Fe (CN) ₆ ] ₂ synthesized, 0.2 ml of water was added, and 0.090 g (3.4 × 10 ⁻⁴ mol) of oleylamine was added. The mixture was mixed with 2 ml of a toluene solution and stirred for one day to obtain Prussian blue-type metal complex ultrafine particles of the present invention in a dispersion.
Centrifugation was performed, the toluene layer was separated, and ultrafine particles having the target metal composition were separated. The absorption spectrum of each of the obtained Prussian blue-type metal complex ultrafine particles was measured.
FIG. 9 shows the result. In FIG. 9, curve 91 shows the result of Prussian blue-type metal complex ultrafine particles with x = 0.8 in the above chemical structural formula, curve 92 shows the result of x = 0.6, and curve 93 shows x = 0. .4 results are shown, and curve 94 shows the results for x = 0.2.

From this UV-visible absorption spectrum, it can be seen that the wavelength of the Fe-Fe charge transfer absorption band is systematically shifted to the longer wavelength side depending on the Ni content. In addition, the intensity of the absorption band derived from Fe—CN—Ni systematically increased around 400 nm. From this result, it is shown that Ni and Fe are uniformly distributed in one nanocrystal (ultrafine particle) (Ni and Fe are inferior to the obtained ultrafine particle (powder)). If it is uniformly distributed in one crystal, or it is simply a mixture of each of the ultrafine particles of Ni ₃ [Fe (CN) ₆ ] ₂ and Fe ₄ [Fe (CN) ₆ ] ₃ In some cases, no systematic shift is observed in the wavelength of the Fe-Fe charge transfer absorption band).

(Preparation Example 5)
Next, samples 1 to 16 were prepared in the same manner as in Preparation Example 1 except that metal atoms M ₁ and M ₂ , ligand L and the dispersion medium were changed as shown in the table below. In the same manner as above, Prussian blue-type metal complex ultrafine particles were prepared, and a dispersion liquid dispersed in a dispersion medium described in the table was obtained. Sample 20 was ultrafine particles obtained in Preparation Example 3, Sample 21 was ultrafine particles obtained in Preparation Example 4, Sample 22 was prepared in the same manner except that oleylamine in Preparation Example 1 was replaced with butyl acetate. Indicates. As can be seen from the results, various Prussian blue-type metal complex ultrafine particles can be produced.

  In all the production examples in the table, the complex crystals synthesized in the step (A) were almost entirely converted into complex fine particles. That is, the yield of ultrafine particles can be reduced to almost 100% by adjusting the charging ratio so that the material for synthesizing the complex crystal is not excessive or insufficient.

In the step of producing the Prussian blue-type metal complex crystal in the step (A), not only the above-described one but also other raw material compounds can be used. For example, in the case of a Prussian _{blue, (NH 4) 4 [Fe} (CN) 6] and is not limited to mixing with _{Fe (NO 3) 3 · 9H} 2 O, K 3 [Fe (CN) 6] Water 1.0g An aqueous solution in which 0.84 g of FeSO ₄ · 7H ₂ O is dissolved in water, or an aqueous solution in which 1.0 g of Na ₄ [Fe (CN) ₆ ] · 10H ₂ O is dissolved in water and Fe ( Prussian blue-type metal complex ultrafine particles to obtain the objective as well by the _{NO 3) 3 · 9H 2 O} 0.83g mixing an aqueous solution dissolved in water. Further, as shown so far, the metal atoms M ₁ and M ₂ are not limited to Fe, and FIG. 10 shows transmission type of cobalt iron cyano complex ultrafine particles manufactured from [Fe (CN) ₆ ] ³⁻ and Co ^2+. An electron microscope image is shown.

  Prussian blue-type metal complex ultrafine particle dispersion (FeHCF-OA) prepared in Preparation Example 1 and Sample No. FIG. 11 shows the results of particle size distribution measurement (using Nikkiso Microtrac UPA-EX150) for 16 Prussian blue-type metal complex ultrafine particle dispersions (NiHCF-OA). The peak of the particle size is present at 30 to 40 nm and is larger than the particle size obtained with an electron microscope or the like. This is because a plurality of particles aggregate and move in the solvent, and indicates the particle size of the secondary particles.

( Reference Example 1)
Sample No. in Table 1 6 and specifically, FeHCF-OA ultrafine particles (Prussian blue-type metal complex ultrafine particles composed of M ₁ = Fe, M ₂ = Fe, L = oleylamine are hereinafter referred to as “FeHCF-OA ultrafine particles”). A dispersion in which 104 mg was dispersed in 4 mg of toluene was prepared. The dispersion glass substrate ITO film by spin coating with a film at room temperature on (vertically 25 mm, lateral 25 mm, rectangular glass substrate having a thickness of 1mm) and substance-adsorbing film layer, sensor of the reference example An electrode body was produced. At this time, spin coating was performed by dropping the dispersion onto the substrate, rotating at a rotation speed of 400 rpm for 30 seconds, and then rotating at 1000 rpm for 10 seconds.
The film thickness of the substance adsorbing thin film layer was measured using a stylus type film thickness measuring machine. In FIG. 12, the portion where the distance d <250 μm is a portion where the material adsorption thin film layer exists, and the portion where d> 250 μm is a portion where the thin film layer is removed. From this, the film thickness of a thin film layer is about 250 nm, and the location dependence of the film thickness is only 10-20 nm, and it turns out that the favorable thin film layer was obtained.

(Example 1 )
Using a dispersion liquid (ink) in which 10 mg of powder of Prussian blue-type metal complex ultrafine particles (M ₁ = Fe, M ₂ = Co) of Table 14 in Table 1 was dispersed in 20 ml of chloroform, the film was formed at room temperature by a chemical bonding film forming method. A material adsorbing thin film layer was formed on the ITO-coated glass substrate (rectangular glass substrate having a length of 10 mm, a width of 40 mm, and a thickness of 1 mm) to produce the sensor electrode body of the present invention. Specifically, the ITO substrate was immersed in a 3- (2-aminoethylaminopropyl) -trimethoxysilane / ethanol solution having a concentration of 3% by mass for 15 minutes, then washed with ethanol, and heated at 110 ° C. for 15 minutes in a heating furnace. Heated and then dried. The obtained substrate was immersed in the dispersion (ink) for 6 hours and then washed with chloroform.

Next, a sensing system configured as shown in FIG. Specifically, changes in electrochemical characteristics with respect to the presence or absence of hydrogen peroxide were measured using the sensor electrode body 133 (glass substrate 133a, ITO conductive film 133b, substance adsorption thin film layer 133c). At this time, Ag / AgCl is used for the reference electrode 134, a platinum electrode is used for the counter electrode 138, and the electrolyte solution 131 is an aqueous phosphate buffer solution (0.05M potassium phosphate dihydrate, 0.05M dipotassium hydrogen phosphate, And 0.10 M potassium nitrate). Each electrode was connected by a terminal so as to perform cyclic voltammetry measurement. 3 mM H ₂ O ₂ was used as a detection target substance.
FIG. 14 shows the result of the cyclic voltammetry measurement. In the cyclic voltammetry 144 of the electrode body for a sensor formed by the chemical bonding film forming method, E0 ′ (Fe ^II / Fe ^III ) = 0.44 V (in 1 M KCl) is shown, and the electrochemical reaction of the electrode fixed species As a result, the peak current value was proportional to the sweep speed.
This sensor electrode body showed electrochemical activity for the redox reaction of hydrogen peroxide (cyclic voltammetry 141 in the figure). When 3 mM hydrogen peroxide was added, the current value at a potential of 1 V was observed to be about 70 times higher than when no hydrogen peroxide was added.

On the other hand, similarly the ITO electrode substrate was not provided with material adsorbed film layer, the measured cyclic voltammetry 142 when adding a cyclic voltammetry 143 and H ₂ O ₂ in H ₂ O ₂ was not added However, H ₂ O ₂ could not be detected.

(Example 2 )
In place of the sample 14 in Table 1, sample 6 (Prussian blue type metal complex ultrafine particles (M ₁ = Fe, M ₂ = Fe)) was used in the same manner as in Example 1, and the sensor electrode body of the present invention. And the sensing system was produced and the change of the electrochemical property was measured. The results are shown in FIG.
This sensor electrode body showed electrochemical activity for the oxidation-reduction reaction of hydrogen peroxide (cyclic voltammetry 151 in the figure). When 3 mM of hydrogen peroxide was added, the current value at a potential of −0.3 V was observed to be about 400 times that of when no hydrogen peroxide was added (cyclic voltammetry 152 in the figure).

( Reference Example 2 )
Sample No. in Table 1 1 dispersion, specifically, 104 mg of FeHCF-OA ultrafine particle powder was dispersed in 4 mg of toluene to prepare a dispersion. Subsequently, the dispersion liquid prepared above was applied to a heart-shaped convex rubber support having a size inscribed in a circle having a diameter of about 8 mm, and pressed and adhered onto a glass substrate coated with ITO. As shown in FIG. 16, a substance adsorbing thin film layer having the same shape as that of the support could be produced on the substrate. Thereby, it turns out that the electrode body for sensors which has a substance-adsorption thin film layer of a desired shape can be simply produced with a letterpress film forming method.

(Example 3 )
FeHCF-OA ultrafine particles were synthesized by the same stirring extraction method as in Preparation Example 1 using iron chloride hexahydrate and sodium ferrocyanide decahydrate as raw materials, and 0.0676 g of the ultrafine particles was added to 1 ml of toluene. An ultrafine particle dispersion was obtained by dispersing. Using the ultrafine particle dispersion, spin-coating (500 rpm for 10 seconds, then 1000 rpm for 30 seconds) on the ITO glass substrate in the same manner as in Reference Example 1 to form a substance-adsorbing thin film layer, thereby producing the sensor electrode body of the present invention. did. This was allowed to stand in acetone for 10 minutes and washed.

In order to confirm the state of the ultrafine particles at this time, infrared spectroscopic measurements were performed before and after the acetone cleaning treatment for the above-described FeHCF-OA fine particles dropped onto KBr pellets. The result is shown in FIG. At this time, after correcting the origin to the measured value of 4000 cm ⁻¹ , normalization was performed with a peak due to CN expansion and contraction. The peak near 2900 cm ⁻¹ is attributed to CH stretching of oleylamine, which is a protective molecule, and the peak near 2080 cm ⁻¹ is attributed to CN stretching of Prussian blue. It can be seen that the peak intensity of CH stretching of oleylamine is reduced by the acetone washing treatment compared with the peak intensity of CN stretching of Prussian blue.

  Cyclic voltammetry measurement was performed on the transparent electrode side member having the ultrafine particles washed with acetone. As a result, the sample before the treatment showed almost no electrochemical response at a voltage of −1.0 V or less, but the one subjected to the acetone treatment showed an electrochemical response (see FIG. 18). From this result, it can be seen that the acetone cleaning treatment improves the electrochemical response of the sensor.

(Example 4 )
FeHCF-OA ultrafine particles were synthesized by the same stirring extraction method as in Preparation Example 1 using iron chloride hexahydrate and sodium ferrocyanide decahydrate as raw materials, and 0.0676 g of the ultrafine particles was added to 1 ml of toluene. An ultrafine particle dispersion was obtained by dispersing. Using the ultrafine particle dispersion, spin coating (500 rpm for 10 seconds, 1000 rpm for 30 seconds) was performed on the ITO glass substrate in the same manner as in Reference Example 1 to form a substance-adsorbing thin film layer, thereby producing the sensor electrode body of the present invention. . This sensor electrode body was left to stand at high temperature (50 ° C., 100 ° C., 150 ° C.) for 2 hours and subjected to heat treatment.

In order to confirm the state of the ultrafine particles at this time, a KBr sample produced in the same manner as in Example 3 was heat-treated at 100 ° C. or 150 ° C., and infrared spectroscopic measurement was performed. The result at this time is also shown in FIG. It can be seen that the relative peak intensity of CH stretching of oleylamine is decreased more than in the case of acetone treatment.
FIG. 19 shows the result of cyclic voltammetry measurement using the transparent electrode side member. For those that were allowed to stand at 150 ° C., the electrochemical response was clearly improved (solid line in the figure). Electrochemical responsiveness was improved even in the case of treatment at 100 ° C. (broken line in the figure). On the other hand, what was processed at 50 degreeC did not improve electrochemical responsiveness (a dashed-dotted line in a figure). From this result, it can be seen that the electrochemical response of the sensor can be improved by performing a heat treatment at a predetermined temperature.

It is sectional drawing which shows typically an example of the preferable aspect of the electrode body for sensors of this invention. It is explanatory drawing which shows typically the structure of the Prussian blue type metal complex ultrafine particle used for the electrode body for sensors of this invention. It is a figure which shows the result of the powder X-ray diffraction of a Prussian blue crystal | crystallization and its ultrafine particle, (I) shows the peak position of a standard sample, (II) shows the measurement result of the obtained ultrafine particle. It is a figure which shows the FT-IR measurement result of Prussian blue crystal | crystallization and its ultrafine particle. It is an ultraviolet visible absorption spectrum of Prussian blue ultrafine particle dispersion. It is a drawing-substituting photograph of a transmission electron microscope image of organic solvent-dispersed Prussian blue ultrafine particles. It is a drawing-substituting photograph of a transmission electron microscope image of water-dispersed Prussian blue ultrafine particles. It is a UV-vis spectrum of cobalt iron cyano complex ultrafine particles (solvent: toluene). It is an absorption spectrum of Prussian blue-type metal complex ultrafine particles obtained by changing the composition of the metal atom M ₂ (iron-nickel composition). It is a drawing substitute photograph of the transmission electron microscope image of cobalt iron cyano complex ultrafine particles. It is a graph which shows the particle size distribution measurement result of the Prussian blue type | mold metal complex ultrafine particle in a dispersion liquid. It is a graph which shows the film thickness measurement result of the substance adsorption thin film layer of the electrode body for sensors of this invention. It is sectional drawing which shows typically the aspect of the sensing system provided with the electrode body for sensors of this invention. It is a measurement result of the cyclic voltammetry which shows the sensor response of hydrogen peroxide in the electrode body for sensors of the present invention. It is a measurement result of the cyclic voltammetry which shows the sensor response of hydrogen peroxide in another electrode body for sensors of the present invention. FIG. 5 is a drawing-substituting photograph showing a sensor electrode body of the present invention in which a Prussian blue-type metal complex ultrafine particle thin film is formed into a heart shape on a relief printing. It is an infrared spectroscopy spectrum when the sample which dripped the FeHCF-OA fine particle dispersion liquid on the KBr pellet was subjected to heating and washing treatment. It is the result of having measured the change of cyclic voltammetry when performing the washing process to the electrode body for sensors of the present invention. It is the result of having measured the change of cyclic voltammetry when heat-processing to the electrode body for sensors of the present invention. It is explanatory drawing which shows typically the crystal structure of a Prussian blue type metal complex.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Material adsorption thin film layer 2 Conductive structure 21 Prussian blue type metal complex (microcrystal)
22 Ligand L
31 X-ray diffraction measurement results of Prussian blue crystals 32 X-ray diffraction measurement results of water-dispersed Prussian blue ultrafine particles 33 X-ray diffraction measurement results of organic solvent-dispersed Prussian blue ultrafine particles 41 Infrared absorption spectrum of Prussian blue crystals 42 Water dispersion Infrared Absorption Spectra of Ultra-Prussian Blue Ultrafine Particles 43 Infrared Absorption Spectra of Organic Solvent-Dispersed Prussian Blue Ultrafine Particles 51 UV-Visible Absorption Spectrum of Prussian Blue Ultrafine Particles (Toluene Dispersion) 52 Absorption spectrum 220 Prussian blue type metal complex 221 Metal atom M ₁
222 carbon atom 223 nitrogen atom 224 metal atom M ₂

Claims (20)

A conductive structure is provided with a substance-adsorbing thin film layer having a thickness of 1 × 10 ⁻⁹ to 1 × 10 ⁻⁶ m formed by a dispersion containing Prussian blue-type metal complex ultrafine particles having a protective ligand. Sensor electrode body,
The sensor applies a voltage to the conductive structure to electrically control the electrode body, and measures a selective change in electrical characteristics when a target substance is adsorbed on the surface of the substance adsorption thin film layer. The target substance is detected by
A compound containing a pyridyl group or an amino group as the protective ligand is added to the Prussian blue type metal complex crystal having the following metal atom M ₁ and metal atom M ₂ obtained by stirring extraction method. Ultrafine particles having an average particle diameter of 200 nm or less coordinated with one or more kinds,
Further, the protective ligand is removed by performing a heat treatment and / or a cleaning treatment with a cleaning agent at the time of forming the substance adsorbing thin film layer and / or thereafter .
[Metal atom M ₁ : At least one metal atom selected from vanadium, chromium, molybdenum, tungsten, manganese, iron, ruthenium, cobalt, nickel, platinum, rhodium, osmium, iridium, palladium, and copper. ]
[Metal atom M ₂ : at least selected from vanadium, chromium, manganese, iron, ruthenium, cobalt, rhodium, nickel, palladium, platinum, copper, silver, zinc, lanthanum, europium, gadolinium, lutetium, barium, strontium, and calcium One metal atom. ]   The sensor electrode body according to claim 1, wherein the target substance is a molecule.   3. The sensor electrode body according to claim 1, wherein the target substance is hydrogen peroxide.   The sensor electrode body according to any one of claims 1 to 3, wherein the substance adsorption thin film layer contains an electrochemical property control agent.   The said substance adsorption thin film layer is a multilayer thin film layer which has at least the thin film layer formed with the said dispersion liquid, and the thin film layer containing the electrochemical characteristic control agent, The any one of Claims 1-4 characterized by the above-mentioned. 2. The electrode body for a sensor according to item 1. The sensor electrode body according to claim 1, wherein the substance-adsorbing thin film layer does not contain a reverse micelle agent .   7. The sensor electrode body according to claim 1, wherein the compound as the protective ligand has 4 to 100 carbon atoms. The sensor electrode body according to any one of claims 1 to 7, wherein the compound as the protective ligand is represented by any one of the following general formulas (1) to (3).
(In the formula, R ₁ and R ₂ each independently represent a hydrogen atom or an alkyl group, alkenyl group, or alkynyl group having 8 or more carbon atoms.)
(In the formula, R ₃ represents an alkyl group, alkenyl group, or alkynyl group having 8 or more carbon atoms.)
(In the formula, R ₄ represents an alkyl group, alkenyl group, or alkynyl group having 6 or more carbon atoms, and R ₅ represents an alkyl group, alkenyl group, or alkynyl group.) The sensor electrode body according to claim 8, wherein the substituents R _{1 to} R ₄ are alkenyl groups.   10. The alkali resistance is increased by containing the protective ligand compound in the mass adsorbing thin film layer in a range of 10 times or less of the Prussian blue type metal complex by mass ratio. The electrode body for sensors according to claim 1.   The substance adsorbing thin film layer is formed by coating the dispersion liquid by any one of a spin coat film forming method, a Langmuir blowet film forming method, a spray film forming method, a layer-by-layer film forming method, and a printing method. The sensor electrode body according to claim 1, which is a liquid film-forming layer having a surface area adsorption surface.   The sensor electrode body according to any one of claims 1 to 11, wherein the dispersion liquid is a dispersion liquid of Prussian blue-type metal complex ultrafine particles prepared by a stirring extraction method. The sensor electrode body according to any one of claims 1 to 12, wherein the stirring extraction method includes the following steps (A) and (B) .
[Step (A): An aqueous solution containing a metal cyano complex (anion) having the metal atom M ₁ as a central metal and an aqueous solution containing a metal cation of the metal atom M ₂ are mixed, and the metal atom M ₁ and step of precipitating crystals of Prussian blue-type metal complex having a metal atom M _2. ]
[Step (B): A step of mixing the solution obtained by dissolving the protective ligand in a solvent with the Prussian blue-type metal complex crystal prepared in Step (A). ]   The sensor provided with the electrode body for sensors of any one of Claims 1-13.   The sensor for food inspection according to claim 14, wherein a trace amount of hydrogen peroxide in food is detected.   The biosensor according to claim 14, wherein the biosensor detects a biological substance.   A sensing system that detects a target substance in a liquid phase by combining the sensor electrode body according to any one of claims 1 to 13, a counter electrode, and a reference electrode. In manufacturing the electrode body according to any one of claims 1 to 13,
More stirring extraction method, a protective ligand comprising a compound containing an amino group or pyridyl group, were dispersed ultrafine particles Prussian blue-type metal complex having a following metal atom M ₁ and metal atom M ₂ Prepare a dispersion,
A material adsorbing thin film layer having a thickness of 1 × 10 ⁻⁹ to 1 × 10 ⁻⁶ m is formed on the conductive structure using the dispersion liquid,
A method for producing an electrode body for a sensor, wherein the protective ligand is removed during and / or after the formation of the substance-adsorbing thin film layer by performing a cleaning treatment with a cleaning agent and / or a heat treatment .
[Metal atom M ₁ : At least one metal atom selected from vanadium, chromium, molybdenum, tungsten, manganese, iron, ruthenium, cobalt, nickel, platinum, rhodium, osmium, iridium, palladium, and copper. ]
[Metal atom M ₂ : at least selected from vanadium, chromium, manganese, iron, ruthenium, cobalt, rhodium, nickel, palladium, platinum, copper, silver, zinc, lanthanum, europium, gadolinium, lutetium, barium, strontium, and calcium One metal atom. ] Scan Pinkoto casting method, the Langmuir-Blodgett film-forming method, and forming a substance-adsorbing thin layer by applying the dispersion liquid by layer-by-layer dip film forming method, and film forming method selected from printing The manufacturing method of the electrode body for sensors of Claim 18. The said stirring extraction method includes the following process (A) and (B), The manufacturing method of the electrode body for sensors of Claim 18 or 19 characterized by the above-mentioned.
[Step (A): An aqueous solution containing a metal cyano complex (anion) having the metal atom M ₁ as a central metal and an aqueous solution containing a metal cation of the metal atom M ₂ are mixed, and the metal atom M ₁ and step of precipitating crystals of Prussian blue-type metal complex having a metal atom M _2. ]
[Step (B): A step of mixing the solution obtained by dissolving the protective ligand in a solvent with the Prussian blue-type metal complex crystal prepared in Step (A). ]