JP2021061240A - Novel mediator - Google Patents

Abstract

To provide an electron mediator which can be used simply and conveniently without need for a complicated process for fixation to an electrode.SOLUTION: The present invention relates to a novel mediator, an electrode modification agent including the mediator, an electron transfer accelerator, and an electrode, a battery, a composition and an enzyme sensor, each including the electron transfer accelerator, and methods for using them. The electrode modification agent comprises a compound having a property of sticking to an electrode neither being coupled with a polymer nor polymerizing, and having a structure given by a formula I below.[In the formula, R1 is a hydrogen atom, a C1-6 alkyl group, an aryl which may be substituted with a sulfo group, or -C(=O)-C1-6 alkyl].SELECTED DRAWING: None

Description

The present invention relates to the use of an aminoanthraquinone-based compound as a mediator, an electrode modifier containing the compound, an electrode containing the compound, an enzyme sensor containing the electrode, and a battery. The present invention also relates to electrochemical measurements using aminoanthraquinone compounds and oxidoreductases, compositions containing aminoanthraquinone compounds and oxidoreductases, and electrodes containing aminoanthraquinone compounds.

In the enzyme electrode, the mediator is used as a mediator for transferring the electrons generated by the redox reaction by the enzyme to the electrode. For efficient electron transfer from the enzyme to the electrode, it is desirable that the mediator is localized near the electrode. Therefore, various methods for immobilizing the mediator on the electrode, such as a method of chemically bonding the electrode and the mediator and a method of polymerizing the mediator itself, are used. However, the conventional method lacks versatility, as the chemical bonding method has restrictions on the side chain functional groups of the mediator, and the method of polymerizing the mediator itself may change the redox potential of the mediator. .. It has also been reported that treating a glassy carbon electrode with a strong acid activates functional groups on the surface to adsorb and fix mediators such as quinones, but it has hardly been put into practical use in the industrial world.

Further, each method requires a complicated process for immobilizing the mediator on the electrode, and has a problem of high cost. Therefore, an electronic mediator that can be easily used without requiring a complicated process for immobilization on an electrode has been desired.

Patent Document 1 (International Publication No. 2011/111531) discloses a device for detecting an analytical substance, and specifically, anthraquinone-based compound, phenanthrenequinone-based compound, and benzoquinone-based compound as analytical substance-sensitive materials. A pH sensor using a compound, an azobenzene-based compound, or the like is described.

International Publication No. 2011/111531 Pamphlet (Patent No. 6049454)

An object of the present invention is to provide a novel mediator that can solve the above problems at least in part.

As a result of diligent efforts to solve the above problems, the present inventors have found that an aminoanthraquinone-based compound can be surprisingly adsorbed on the electrode surface, and that the compound can function as a mediator. , The present invention has been completed. To the best of our knowledge, it has been reported so far that 1-acetamido-4-hydroxyanthraquinone, 1-amino-4-hydroxyanthraquinone, and 2-amino-3-hydroxyanthraquinone are mediators that adsorb to the electrode. Not, this is a surprising finding.

［式中、R¹は、水素、C_1-6アルキル、場合によりスルホにより置換されてもよいアリール又は-C(=O)-C_1-6アルキルである］
を有する化合物を含む、電極改変剤。
[２] ポリマーに結合されることなく、若しくはポリマー化することなく電極に吸着する性質を有し、式Iの構造、

［式中、R¹は、水素、C_1-6アルキル、場合によりスルホにより置換されてもよいアリール又は-C(=O)-C_1-6アルキルである］
を有する化合物を含む、電子伝達促進剤。
[３] 実施形態１に記載の電極改変剤又は実施形態２に記載の電子伝達促進剤を含む、電池。
[４] 前記化合物が前記電池の電極に固定化されている、実施形態３に記載の電池。
[５] 酸化還元酵素を含む、実施形態３又は４に記載の電池。
[６] 酸化還元酵素が電極に固定化されている、実施形態５に記載の電池。
[７] 実施形態１に記載の電極改変剤又は実施形態２に記載の電子伝達促進剤を含む、組成物。
[８] 実施形態１に記載の電極改変剤又は実施形態２に記載の電子伝達促進剤を含む、電極。
[９] 酵素を有する、実施形態８に記載の電極、又は、酵素を含む、実施形態７に記載の組成物。
[１０] 酵素が酸化還元酵素である、実施形態９に記載の電極又は組成物。
[１１] 酸化還元酵素が固定化されている、実施形態１０に記載の電極。
[１２] 実施形態１に記載の電極改変剤又は実施形態２に記載の電子伝達促進剤を含むセンサー、又は、実施形態１０若しくは１１に記載の電極を有する酵素センサー。
[１３] 化合物が、以下の構造

を有する1-アセトアミド-4-ヒドロキシアントラキノン、
以下の構造

を有する1-アミノ-4-ヒドロキシアントラキノン、又は
以下の構造

を有する2-アミノ-3-ヒドロキシアントラキノン、
又はその塩、無水物若しくは溶媒和物である、
実施形態１に記載の電極改変剤、
実施形態２に記載の電子伝達促進剤、
実施形態３〜６のいずれかに記載の電池、
実施形態７、９又は１０に記載の組成物、
実施形態８〜１１のいずれかに記載の電極、又は
実施形態１２に記載の酵素センサー。
[１４] 実施形態１に記載の電極改変剤又は実施形態２に記載の電子伝達促進剤を用いる工程を含む、電池の製造方法。
[１５] 前記電極改変剤、又は電子伝達促進剤を、電池の電極と接触させる工程を含む、実施形態１４に記載の方法。
[１６] 実施形態３〜６及び１３のいずれかに記載の電池を用いる、発電方法。
[１７] 実施形態７、９、１０及び１３のいずれかに記載の組成物、
実施形態８〜１１、及び１３のいずれかに記載の電極、又は
実施形態１２又は１３に記載のセンサー
を用いる、電気化学的測定方法。
[１８] 実施形態１又は１３に記載の電極改変剤、
実施形態２又は１３に記載の電子伝達促進剤、又は
実施形態７、９、１０及び１３のいずれかに記載の組成物、
を電極に接触させる工程を含む、電極を改変又は修飾する方法。 The present disclosure includes the following embodiments:
[1] The structure of the formula I, which has the property of adsorbing to the electrode without being bonded to the polymer or polymerized.

[In the formula, R ¹ is hydrogen, C _1-6 alkyl, optionally aryl or -C (= O) -C _1-6 alkyl optionally substituted with sulfo].
An electrode modifier comprising a compound having.
[2] The structure of the formula I, which has the property of adsorbing to the electrode without being bonded to the polymer or polymerized.

[In the formula, R ¹ is hydrogen, C _1-6 alkyl, optionally aryl or -C (= O) -C _1-6 alkyl optionally substituted with sulfo].
An electron transfer promoter, which comprises a compound having.
[3] A battery comprising the electrode modifier according to the first embodiment or the electron transfer promoter according to the second embodiment.
[4] The battery according to the third embodiment, wherein the compound is immobilized on an electrode of the battery.
[5] The battery according to embodiment 3 or 4, which comprises a redox enzyme.
[6] The battery according to the fifth embodiment, wherein the redox enzyme is immobilized on an electrode.
[7] A composition comprising the electrode modifier according to the first embodiment or the electron transfer promoter according to the second embodiment.
[8] An electrode containing the electrode modifier according to the first embodiment or the electron transfer promoter according to the second embodiment.
[9] The electrode according to embodiment 8 having an enzyme, or the composition according to embodiment 7 containing an enzyme.
[10] The electrode or composition according to embodiment 9, wherein the enzyme is an oxidoreductase.
[11] The electrode according to embodiment 10, wherein the redox enzyme is immobilized.
[12] A sensor containing the electrode modifier according to the first embodiment or the electron transfer promoter according to the second embodiment, or an enzyme sensor having the electrode according to the tenth or eleventh embodiment.
[13] The compound has the following structure

Has 1-acetamido-4-hydroxyanthraquinone,
The following structure

1-Amino-4-hydroxyanthraquinone with, or the following structure

Has 2-amino-3-hydroxyanthraquinone,
Or its salt, anhydride or solvate,
The electrode modifier according to the first embodiment,
The electron transport chain according to the second embodiment,
The battery according to any one of embodiments 3 to 6.
The composition according to embodiment 7, 9 or 10.
The electrode according to any one of embodiments 8 to 11, or the enzyme sensor according to embodiment 12.
[14] A method for manufacturing a battery, which comprises a step of using the electrode modifier according to the first embodiment or the electron transfer accelerator according to the second embodiment.
[15] The method according to embodiment 14, comprising a step of bringing the electrode modifier or the electron transfer accelerator into contact with the electrodes of the battery.
[16] A power generation method using the battery according to any one of embodiments 3 to 6 and 13.
[17] The composition according to any one of embodiments 7, 9, 10 and 13.
An electrochemical measurement method using the electrode according to any one of embodiments 8 to 11 and 13, or the sensor according to embodiment 12 or 13.
[18] The electrode modifier according to embodiment 1 or 13.
The electron transfer promoter according to embodiment 2 or 13, or the composition according to any one of embodiments 7, 9, 10 and 13.
A method of modifying or modifying an electrode, which comprises the step of bringing the electrode into contact with the electrode.

This specification includes the disclosure of Japanese Patent Application No. 2019-183555, which is the basis of the priority of the present application.

Unlike conventional mediators, the aminoanthraquinone-based compound of the present invention can be adsorbed on the electrode surface as it is without requiring special treatment such as acid treatment of the electrode or polymerization of the mediator itself, and is therefore convenient. Can be immobilized on the electrode. Moreover, such an electrode can be used for an electrochemical measurement. Moreover, such an electrode can be applied to a battery.

Cyclic voltammetry using 1-acetamide-4-hydroxyanthraquinone was performed, and the results of plotting the _{sweep rate and I Omax are shown.} Cyclic voltammetry using 1-amino-4-hydroxyanthraquinone was performed, and the results of plotting the _{sweep rate and I Omax are shown.} Cyclic voltammetry using 2-amino-3-hydroxyanthraquinone was performed, and the results of plotting the _{sweep rate and I Omax are shown.} Cyclic voltammetry using p-phenylenediamine was performed, and the results of plotting the _{sweep rate and I Omax are shown.}

(Amino anthraquinone compound)
(Electrodes on which aminoanthraquinone compounds are adsorbed)
In certain embodiments, the present invention provides aminoanthraquinone compounds. In another embodiment, the present invention provides an electrode modifier comprising an aminoanthraquinone compound. This electrode modifier can be adsorbed on the surface of the electrode and can modify the electron acceptability of the electrode. In another embodiment, the present invention provides an electrode on which an aminoanthraquinone compound or an electrode modifier of the present invention is adsorbed. In another embodiment, the present invention provides an electrochemical measurement composition containing an aminoanthraquinone-based compound or an electrode modifier of the present invention. The composition may further contain an enzyme. The enzyme can be an oxidoreductase. In another embodiment, the present invention provides an electrochemical measurement kit containing an aminoanthraquinone-based compound or an electrode modifier of the present invention. In the present specification, the electrode modifier may be referred to as an electrode adsorbent.

The aminoanthraquinone-based compound of the present invention can be adsorbed on the electrode surface by being brought into contact with the electrode. At this time, no special operation such as acid treatment of the electrode surface to activate it is required in order to adsorb it to the electrode. In a certain embodiment, the property of the compound of the present invention that adsorbs to an electrode means the property that the compound physically adsorbs to the electrode, and the electrode can be made of a material such as carbon, gold, or platinum. Further, the present invention includes an embodiment in which the compound is adsorbed on a primary material such as carbon powder or carbon material, and then the primary material such as carbon powder or carbon material is immobilized on an electrode. However, this is an explanation of the properties of the compound of the present invention, and does not limit the method of using the compound. That is, in certain embodiments, the present invention provides a method of adsorbing a compound of the present invention on a primary material such as carbon powder or carbon material and then immobilizing the primary material such as carbon powder or carbon material on an electrode.

In certain embodiments, the present invention provides a primary material, such as a carbon powder or carbon material adsorbed with a compound of the present invention. The primary material such as this carbon powder or carbon material can be further applied or applied to the electrode. Examples of the primary material include, but are not limited to, carbon, platinum, and gold. Examples of the carbon material include carbon black, carbon fiber, single-walled or multi-walled carbon nanotubes, graphene, and Ketjen black.

In a certain embodiment, the property of the compound of the present invention to adsorb to an electrode, carbon powder or carbon material means the property of the compound to directly and physically adsorb to the electrode, carbon powder or carbon material. It does not refer to the property of covalently bonding to a polymer or linker to bond to an electrode, carbon powder or carbon material. In the present specification, such a property is adsorbed to a carbon powder or a carbon material without being bonded to a polymer or adsorbed to an electrode, or without being bonded to a polymer or polymerizing the compound. There is a property. However, this is an explanation of the properties of the compound of the present invention, and does not limit the method of using the compound. That is, in certain embodiments, the present invention provides a method of covalently bonding a compound of the present invention to a polymer or linker to bond it to an electrode, carbon powder or carbon material. Further, the fact that the compound directly and physically adsorbs to the electrode, carbon powder or carbon material does not prevent the compound of the present invention itself (as a monomer) from covalently bonding to the carbon powder or carbon material. In addition, in this specification, a polymer means a polymer in which the same unit is multimerized by, for example, 10 or more.

In certain embodiments, the carbon powder or carbon material, carbon electrode, gold electrode, and platinum electrode for adsorbing the compound of the present invention are not acid-treated. That is, in a certain embodiment, the acid-treated carbon powder or carbon material and carbon electrode are removed from the carbon powder or carbon material that adsorbs the compound of the present invention.

In certain embodiments, the compounds of the invention can be used in combination with enzymes. In certain embodiments, the compounds of the invention can be used in combination with oxidoreductase. In certain embodiments, the compounds of the present invention can be used as electron transfer promoters. In certain embodiments, the aminoanthraquinone compounds of the invention function as mediators in redox reactions catalyzed by oxidoreductases. Examples of oxidoreductases include various oxidoreductases classified into EC Group 1, such as glucose oxidase, glucose dehydrogenase, amadriase (also referred to as fructosyl peptide oxidase or fructosyl amino acid oxidase), peroxidase, galactose oxidase, and bilirubin oxidase. Pyruvate oxidase, D- or L-amino acid oxidase, amine oxidase, cholesterol oxidase, choline oxidase, xanthin oxidase, sarcosine oxidase, D- or L-lactate oxidase (LOD), ascorbic acid oxidase, cytochrome oxidase, alcohol dehydrogenase, cholesterol dehydrogenase , Oxidoreductase, Oxidoreductase, Fructose Dehydrogenase (FDH), Sorbitol Dehydrogenase, D- or L-Lactate Dehydrogenase, Appleic Acid Dehydrogenase, Gloxy Dehydrogenase, 17B Hydroxysteroid Dehydrogenase, Estradiol 17B Dehydrogenase, D- or L-Amino Acid Dehydrogenase Alaldehyde 3-phosphate dehydrogenase, 3-hydroxysteroid dehydrogenase, diaholase, catalase, glutathione reductase, titochrome b5 reductase, adrenoxin reductase, titochrome b5 reductase, adrenodoxine reductase, nitrate reductase, phosphate dehydrogenase, bilirubin oxidase, lacquer Examples thereof include, but are not limited to, polyamine oxidase, formate dehydrogenase, pyranose oxidase, pyranose dehydrogenase, and tauropine dehydrogenase. Moreover, it may be a combination of a plurality of enzymes. Examples of the coenzyme of the above enzyme include nicotinamide adenine dinucleotide (NAD), nicotinamide adenine dinucleotide phosphate, flavin adenine dinucleotide (FAD), and pyrroloquinoline quinone. That is, examples of glucose dehydrogenase (GDH) include FAD-dependent GDH, NAD-dependent GDH, and PQQ-dependent GDH. The activities of the redox enzymes listed above can be measured using various substrates by, for example, the methods described in Methods in Enzymology (Volumes 1 to 602).

In the present specification, the function as a mediator means that an aminoanthraquinone-based compound contributes to electron transfer. For example, in a system using an electrode, the aminoanthraquinone-based compound of the present invention receives an electron from an oxidoreductase and reduces it. It becomes a mold and passes electrons to the electrode to return to the oxidized mold. From this point of view, the aminoanthraquinone-based compound of the present invention that functions as a mediator can also be called an electron transfer mediator, an electron transfer promoter, or an electron mediator (also simply referred to as a mediator). These terms are synonymous herein.

In certain embodiments, electrodes having acid-treated carbon powder or carbon particles are excluded from the electrodes of the present invention.

In certain embodiments, the redox enzyme used with the electrodes of the present invention may be immobilized on the electrodes. That is, in this embodiment, the present invention provides an electrode in which an oxidoreductase is immobilized and an electrode modifier of the present invention is adsorbed. In certain embodiments, the present invention provides an enzyme sensor comprising an electrode in which an oxidoreductase is immobilized and an electrode modifier of the present invention is adsorbed. In certain embodiments, a sensor comprising the electrode modifier or electron transfer promoter of the present invention is provided.

The aminoanthraquinone-based compound of the present invention or an electrode modifier, an electrode adsorbent or an electron transfer promoter containing the aminoanthraquinone-based compound can be adsorbed on the electrode surface without requiring special treatment. Therefore, in a certain embodiment, the aminoanthraquinone-based compound of the present invention, the electrode modifying agent, the electrode adsorbent, or the electron transfer promoter may be adsorbed on the electrode in advance in order to prepare the modified electrode. In another embodiment, when performing an electrochemical measurement, a measurement solution (composition) containing the aminoanthraquinone compound of the present invention, an electrode modifier, an electrode adsorbent or an electron transfer promoter, and an oxidoreductase is used. Can be used. In this embodiment, the aminoanthraquinone of the present invention is obtained by physically contacting the electrode with a composition containing the aminoanthraquinone-based compound of the present invention, an electrode modifier, an electrode adsorbent or an electron transfer promoter, and an oxidative reduction enzyme. An electrode in which a system compound, an electrode modifier, an electrode adsorbent or an electron transfer promoter is adsorbed on the electrode and the electrochemical properties are modified is produced on the spot at the time of measurement.

In certain embodiments, the electrodes are not pretreated with an acid when the aminoanthraquinone compounds, electrode modifiers, electrode adsorbents or electron transfer promoters of the present invention are adsorbed on the electrodes. In another embodiment, the electrode may be pretreated with an acid in adsorbing the aminoanthraquinone compound, the electrode modifier, the electrode adsorbent or the electron transfer promoter of the present invention on the electrode. The pretreatment with an acid referred to here includes bringing an electrolyte solution containing an acid into contact with the electrode. Further, in the present specification, the acid means an acid having a pH of 4 or less, for example, pH 4 or less, pH 3 or less, pH 3 or less, pH 2 or less, pH 2 or less, and pH 1. Further, in a certain embodiment, the aminoanthraquinone compound, the electrode modifier, the electrode adsorbent or the electron transfer promoter of the present invention is converted into an oxidized form or a reduced form by applying a specific potential and then adsorbed on the electrode. Is also good. An oxidizing agent or a reducing agent may be used as a method for converting to the oxidized form or the reduced form, and the method is not particularly limited. Further, in a certain embodiment, the aminoanthraquinone compound of the present invention, an electrode modifier, an electrode adsorbent or an electron transfer promoter may be encapsulated in a polymer and adsorbed on the electrode. Such adsorption is referred to herein as embedded or entrapped. This is distinguished from adsorption fixation, that is, fixation by physical adsorption. In inclusion fixation or inclusion fixation, the compound of the present invention can be diffused in the polymer, whereas in adsorption fixation, the compound of the present invention does not diffuse or hardly diffuses. In another embodiment, the compounds of the present invention, electrode modifiers, electrode adsorbents or electron transfer promoters are used in ionic polymers such as cationic polymers such as polyethyleneimine and polylysine, and anionic polymers such as polyaniline and polyacrylic acid. It may be adsorbed on the electrode by electrostatic interaction using or the like. In another embodiment, the compound of the present invention, an electrode modifier, an electrode adsorbent or an electron transfer promoter may be adsorbed or immobilized on an enzyme with a cross-linking agent and immobilized on the electrode together with the enzyme. In another embodiment, the adsorbed compound of the present invention may be polymerized by a redox reaction or a cross-linking agent. In certain embodiments, the aminoanthraquinone compounds of the present invention may be included in an electrolyte tank or electrolyte solution in contact with the electrodes.

［式中、R¹は、水素、C_1-6アルキル、場合によりスルホにより置換されてもよいアリール又は-C(=O)-C_1-6アルキルである］。R¹が水素である場合はアミノアントラキノンであり、R¹がC_1-6アルキル、場合によりスルホにより置換されてもよいアリール、-C(=O)-C_1-6アルキルであるようにアルキルアミノ基、アミド基、場合によりスルホにより置換されてもよいアリールアミノ基が付加されたアントラキノンである場合にも、本発明のアミノアントラキノン系化合物に含めるものとする。アントラキノン上の他の位置にアミノ基、アルキルアミノ基、アミド基、場合によりスルホにより置換されてもよいアリールアミノ基及びヒドロキシ基を有する構造異性体についても同様である。本明細書において、場合によりある基により置換されてもよい、とは、当該基により置換されている、または置換されていないことをいう。アリール基としては例えばフェニル基、トリル基、キシリル基、ナフチル基が挙げられるがこれに限らない。トリル基はトルエンから誘導されるCH₃C₆H₄-基をいう。スルホ基はSO₃H-基をいう。本明細書においてスルホというときは、その塩、例えばNa塩も含まれるものとする。 In certain embodiments, the aminoanthraquinone compounds of the invention can be compounds of the following General Formula I or salts, anhydrides or solvates thereof:

[In the formula, R ¹ is hydrogen, C _1-6 alkyl, optionally aryl or -C (= O) -C _1-6 alkyl optionally substituted with sulfo]. When R ¹ is hydrogen or an amino anthraquinone, alkyl as R ¹ is a C _1-6 alkyl, optionally may be substituted with sulfo by aryl, -C (= O) -C _1-6 alkyl Anthraquinone to which an amino group, an amide group, and optionally an arylamino group which may be substituted by a sulfo is added is also included in the aminoanthraquinone-based compound of the present invention. The same applies to structural isomers having an amino group, an alkylamino group, an amide group, and optionally an arylamino group and a hydroxy group at other positions on anthraquinone. As used herein, the term "may be substituted by a certain group" means that it is substituted or not substituted by the group. Examples of the aryl group include, but are not limited to, a phenyl group, a tolyl group, a xsilyl group, and a naphthyl group. A tolyl group is a CH ₃ C ₆ H ₄ -group derived from toluene. The sulfo group refers to the SO ₃ H- group. When the term sulfo is used herein, the salt thereof, for example, Na salt is also included.

を有する（CAS番号7323-62-8）。 In certain embodiments, the aminoanthraquinone compounds of the invention can be 1-acetamide-4-hydroxyanthraquinone. 1-Acetamide-4-hydroxyanthraquinone has the following structure

Has (CAS No. 7323-62-8).

を有する（CAS番号116-85-8）。 In another embodiment, the aminoanthraquinone-based compound of the present invention can be 1-amino-4-hydroxyanthraquinone. 1-Amino-4-hydroxyanthraquinone has the following structure

Has (CAS No. 116-85-8).

を有する（CAS番号117-77-1）。別の実施形態において、本発明のアミノアントラキノン系化合物は1-ヒドロキシ-4-トルイジノアントラキノン（CAS番号81-48-1）、又はアシッドバイオレット43（CAS番号4430-18-6）でありうる。なお、2-カルボキシアントラキノン、アセチルアミドアントラキノン、及び2-βナフトール-メチルアントラキノンは、本発明のアミノアントラキノン系化合物に含まれない。2-カルボキシアントラキノン、アセチルアミドアントラキノン、及び2-βナフトール-メチルアントラキノンはヒドロキシル基を有しないため、本発明のアミノアントラキノン系化合物とは化学的な性質が異なると考えられる。 In another embodiment, the aminoanthraquinone compound of the present invention can be 2-amino-3-hydroxyanthraquinone. 2-Amino-3-hydroxyanthraquinone has the following structure

Has (CAS No. 117-77-1). In another embodiment, the aminoanthraquinone compounds of the invention can be 1-hydroxy-4-toluidinoanthraquinone (CAS No. 81-48-1) or Acid Violet 43 (CAS No. 4430-18-6). .. 2-Carboxyanthraquinone, acetylamide anthraquinone, and 2-βnaphthol-methylanthraquinone are not included in the aminoanthraquinone compounds of the present invention. Since 2-carboxyanthraquinone, acetylamide anthraquinone, and 2-βnaphthol-methylanthraquinone do not have a hydroxyl group, they are considered to have different chemical properties from the aminoanthraquinone compounds of the present invention.

As used herein, alkyl refers to a straight-chain or branched-chain hydrocarbon having, for example, 6 carbon atoms. Examples of alkyl include, but are not limited to, methyl, ethyl, propyl, isopropyl, isobutyl, n-butyl, tert-butyl, isopentyl, n-pentyl and heptyl. Examples of -C (= O) -C _1-6 alkyl include, but are not limited to, acetyl, propionyl, butyryl, isobutyryl, valeryl, isovaleryl, and pivaloyl.

As used herein, the number of atoms (such as carbon atoms) is expressed, for example, as "Cx-Cy alkyl", which refers to an alkyl group having x to y carbon atoms. The same notation is used for other substituents and ranges.

As used herein, "optionally substituted" and "substituted or not substituted" mean any substitution with one or more substituents, including multiple substitutions. Is done.

The functional group for covalently bonding the aminoanthraquinone compound of the present invention with the enzyme may be modified. C1 to C20 alkyls, amino acids, peptides, etc. may be inserted as a linker between the aminoanthraquinone compound and the functional group. A hydroxyl group, an amino group, an alkene or the like may be appropriately contained between the linkers. As a functional group, for example, a hydroxyl group, a carboxyl group, an amino group, an aldehyde group, a hydradino group, a thiocyanate group, an epoxy group, a vinyl group, a halogen group, an acid ester group, a phosphoric acid group, a thiol group, a disulfide group and a dithiocarbamate group. , Dithiophosphate group, dithiophosphonate group, thioether group, thiosulfate group, succinimide group, maleimide group, thiourea group and the like.

The aminoanthraquinone-based compound may have a redox state and an ionized state. In the above chemical formula, the aminoanthraquinone-based compound of the present invention is described in a neutral and reduced form. However, not only in this form, the aminoanthraquinone-based compound of the present invention can be in an oxidized form, a semi-oxidized form, or a reduced form. Further, the aminoanthraquinone-based compound of the present invention may be a neutral type or a cationic type. For convenience, when referring to the aminoanthraquinone-based compound of the present invention, for example, the aminoanthraquinone-based compound of the present invention represented by the above chemical formula, this is a neutral or cationic type, an oxidized type, a semi-oxidized type, or a reduced type. It shall include those in the form. For example, after adding a neutral and oxidized compound as the aminoanthraquinone compound of the present invention to the measurement system, it may be changed to an oxidized and cationic compound by the pH of the solution and electron transfer. However, such compounds are also included in the aminoanthraquinone compounds of the present invention. The aminoanthraquinone-based compound of the present invention includes its salts, acid addition salts, anhydrides and solvates. Examples of the salt include, but are not limited to, Group 1 element salts and Group 17 element salts, such as Na salt, K salt, Cl salt, and Br salt. Examples of the acid addition salt include, but are not limited to, hydrochlorides, sulfates, sulfites, and nitrates.

The aminoanthraquinone-based compound of the present invention may be artificially synthesized or a natural product may be obtained. It may also be commercially available. In the case of synthesis, organic synthesis can be performed using a conventional organic synthesis method, and the product can be confirmed by NMR, IR, mass spectrometry, or the like. Unless otherwise specified, the practice of the present invention uses prior arts of chemistry, organic synthesis, biochemistry, molecular biology, and electrochemistry, which are within the ability of those skilled in the art. Such techniques are described in the literature. For example, Organic Chemistry (Jonathan Clayden (edit), Nick Greeves (author), Stuart Warren (author), Peter Wothers (author)), Oxford Univ Pr, 2000 and March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure (Michael). See B. Smith (Author), Jerry March (Author)) Wiley-Interscience, 6th edition, 2007. Each of these general texts is incorporated herein by reference.

(Method of immobilizing enzyme or oxidoreductase)
The enzyme or oxidoreductase can be immobilized on the solid phase by any known method. Enzymes such as oxidoreductase may be immobilized on beads, membranes, carbon particles, gold particles, platinum particles, polymers, or electrode surfaces. Examples of the immobilization method include a method using a cross-linking reagent, a method of encapsulating in a polymer matrix, a method of coating with a dialysis membrane, a photocrosslinkable polymer, a conductive polymer, an oxidation-reduced polymer, and the like. It may be adsorbed and fixed on the top, or these may be used in combination. Typically, oxidoreductase is immobilized on a carbon electrode with glutaraldehyde and then treated with a reagent having an amine group to block glutaraldehyde. The amount of oxidoreductase to be immobilized can be an amount capable of generating a current required for electrochemical measurement or fuel cell power generation, and can be appropriately determined.

(Method for Adsorbing Amino Anthraquinone Compounds of the Present Invention)
In certain embodiments, the aminoanthraquinone compounds of the present invention may be present in a free state in a solution, and may be adsorbed on beads, membranes, carbon particles, gold particles, platinum particles, polymers, or electrode surfaces. For example, it may be physically adsorbed. The fact that the aminoanthraquinone-based compound of the present invention is adsorbed on the electrode surface may be immobilized on the electrode surface, but these are synonymous in the present specification. Examples of the adsorption method include a step of dissolving the aminoanthraquinone compound of the present invention in an appropriate medium and physically contacting the solution with the electrode. In another embodiment, the aminoanthraquinone compounds of the present invention may be sprayed onto the electrodes. The amount of the aminoanthraquinone-based compound to be adsorbed can be an amount capable of generating a current required for electrochemical measurement or battery power generation, and can be appropriately determined.

In certain embodiments, the aminoanthraquinone-based compound of the present invention may be adsorbed on the electrode, and an enzyme such as an oxidoreductase may be immobilized. In this case, the aminoanthraquinone-based compound of the present invention may be first adsorbed and then the enzyme such as oxidoreductase may be immobilized, or the enzyme such as oxidoreductase may be immobilized first and then the aminoanthraquinone-based compound of the present invention may be adsorbed. Alternatively, the aminoanthraquinone-based compound may be adsorbed at the same time in the operation of immobilizing the enzyme, for example, the oxidoreductase.

The final concentration of the aminoanthraquinone compound of the present invention added to the sample solution is not particularly limited, and for example, 1 pM or more, 2 pM or more, 3 pM or more, 4 pM or more, 5 pM or more, 6 pM or more, 7 pM or more, 8 pM or more, 9 pM or more, 10pM or more, 1M or less, 100mM or less, 20mM or less, 10mM or less, 5mM or less, 1mM or less, 800μM or less, 600μM or less, 500μM or less, 400μM or less, 300μM or less, 200μM or less, 100μM or less, 50μM or less, for example, 1pM to 1M, 1pM to 100 mM, 1 pM to 20 mM, 1 pM to 10 mM, 1 pM to 5 mM, 2 pM to 1 mM, 3 pM to 800 μM, 4 pM to 600 μM, 5 pM to 500 μM, 6 pM to 400 μM, 7 pM to 300 μM, 8 pM to 200 μM, 9 pM to 100 μM, 9 pM to 100 μM It can be in the range of 50 μM. The final concentration of the aminoanthraquinone compound of the present invention added to the sample solution is not particularly limited, and is, for example, 0.000001 to 0.5% (w / v), 0.000003 to 0.3% (w / v), 0.000005 to 0.1% (w / v). ), 0.00001 to 0.05% (w / v), 0.00002 to 0.03% (w / v), 0.00003 to 0.01% (w / v). The order of addition of the aminoanthraquinone compound and other reagents is not limited, and they may be added simultaneously or sequentially.

In certain embodiments, the time for redox reaction or electrochemical measurement can be 60 minutes or less, 30 minutes or less, 10 minutes or less, 5 minutes or less, or 1 minute or less. Alternatively, in enzyme sensors and batteries that measure for a long period of time, the time for redox reaction is 60 minutes or more, 120 minutes or more, 1 day or more, 2 days or more, 3 days or more, 1 week or more, 2 weeks or more, It can be 3 weeks or more. For example, when an aminoanthraquinone compound is used with an oxidoreductase, the concentration of each component of the electrochemical measurement reagent is adjusted according to the concentration range of the mediator contained in the sample or estimated to be produced in the sample. be able to.

Unless otherwise specified, the enzyme contained in the composition of the present invention or the enzyme immobilized on the electrode of the present invention is a purified enzyme. Cell extracts containing enzymes, cell crushed solutions, and crude enzyme extracts contain various contaminants in addition to enzymes. For example, in microorganisms, the amount of riboflavin in crude enzyme extract has been reported to be about 53 to 133 μM (J Indust Micro Biotech 1999, 22, pp. 8-18). If such a crude enzyme extract or the like is subjected to an electrochemical measurement as it is, the contaminating substances receive electrons and the transfer of electrons to the electrode is hindered. Accurate electrochemical measurements are difficult. Therefore, in the electrochemical measurement method of the present invention, an enzyme from which contaminants have been removed can be used. As used herein, the term "purified enzyme" or "purified enzyme" means that the protein does not necessarily have to be a pure product, and contaminants have been removed from the enzyme preparation to the extent that it can be measured electrochemically. It means that you are.

(Measurement of redox enzyme activity)
In explaining the measurement of the activity of oxidoreductase, glucose dehydrogenase (GDH) and peroxidase will be taken as specific examples. GDH (EC 1.1.5.9) catalyzes the reaction of oxidizing the hydroxyl groups of glucose to produce glucono-δ-lactone. At this time, the electron acceptor receives an electron and becomes a reduced electron acceptor. Using this principle of action, the activity of GDH can be measured using, for example, the following measurement system using phenazinemethsulfate (PMS) and 2,6-dichloroindophenol (DCIP) as electron acceptors. ..
(Reaction 1) D-glucose + PMS (oxidized type)
→ D-glucono-δ-lactone + PMS (reduced form)
(Reaction 2) PMS (reduced type) + DCIP (oxidized type)
→ PMS (oxidized type) + DCIP (reduced type)

Specifically, first, in (Reaction 1), PMS (reduced form) is produced with the oxidation of D-glucose. As the PMS (reduced form) is oxidized by the subsequent progress (reaction 2), DCIP is reduced. The degree of disappearance of this "DCIP (oxidized type)" can be detected as the amount of change in absorbance at a wavelength of 600 nm, and the enzyme activity can be determined based on this amount of change.

GDH activity can be measured according to the following procedure. Mix 2.05 mL of 100 mM phosphate buffer (pH 7.0), 0.6 mL of 1M D-glucose solution and 0.15 mL of 2 mM DCIP solution and incubate at 37 ° C for 5 minutes. Then 0.1 mL of 15 mM PMS solution and 0.1 mL of enzyme sample solution are added to initiate the reaction. The absorbance at the start of the reaction and over time is measured, the amount of decrease in absorbance at 600 nm per minute (ΔA600) with the progress of the enzymatic reaction is determined, and the GDH activity is calculated according to the following formula. At this time, GDH activity defines 1 U as the amount of enzyme that reduces 1 μmol of DCIP per minute in the presence of D-glucose at a concentration of 200 mM at 37 ° C.

In the formula, 3.0 is the liquid volume of reaction reagent + enzyme reagent (mL), 16.3 is the absorbance of the mmol molecule (cm ^{2 /} μmol) under the present activity measurement conditions, 0.1 is the liquid volume of the enzyme solution (mL), and 1.0 is the liquid volume of the enzyme solution. Cell optical path length (cm), ΔA 600 _blank is the amount of decrease in absorbance at 600 nm per minute when the reaction is started by adding 100 mM phosphate buffer (pH 7.0) instead of the enzyme sample solution, and df is dilution. Represents a multiple.

Peroxidase (EC 1.11.1.7) catalyzes a reaction that cleaves the peroxide structure of a compound. At this time, electrons are extracted from the electron donor and converted into an oxidized electron donor. Using this principle, the activity of peroxidase can be measured using, for example, the following measurement system using hydrogen peroxide as a compound having a peroxide structure and pyrogallol as an electron donor.
2 Pyrogallol + 3H ₂ O ₂
→ Purpurogallin + 5H ₂ O + CO ₂
The enzyme activity can be determined by extracting the produced purpurogallin with ether and measuring the absorbance at 420 nm.

The enzymatic activity of peroxidase can be measured according to the following procedure. Mix 14.0 ml of distilled water, 2.0 ml of 5% (w / v) pyrogallol aqueous solution prepared at the time, 1.0 ml of 0.147 M aqueous hydrogen peroxide solution prepared at the time of use, and 2.0 ml of 0.1 M phosphate buffer (pH 6.0). Then preheat at 20 ° C for about 5 minutes. This is designated as the reaction mixture 1. Subsequently, 1.0 ml of an enzyme solution dissolved in 0.1 M phosphate buffer (pH 6.0) previously ice-cooled is added, and the reaction is started. After reacting at 20 ° C for exactly 20 seconds, add 1.0 ml of 2.0 N sulfuric acid to stop the reaction. Extract purpurogallin produced from the mixed solution after the reaction is stopped with 15 ml of ether. Repeat this operation 5 times, combine the extracts, and add ether to make the total volume 100 ml. Measure the absorbance of this solution at 420 nm (this value is called the OD test).

For blinding, the reaction mixture 1 is left at 20 ° C. for 20 seconds, then 1.0 ML of 2.0 N sulfuric acid is added and mixed, and then 1.0 ML of the enzyme solution is added to prepare the reaction mixture. Ether extraction is performed on this solution in the same manner as above, and the absorbance is measured (this value is referred to as OD BLANK).

From the OD test and OD blank, and the dilution ratio (df) of the enzyme solution, the peroxidase activity is calculated based on the following formula. Under the above conditions, the amount of enzyme that produces 1.0 mg of purpurogallin in 20 seconds is defined as 1 U. In the following formula, 0.117 means the absorbance of a 1 mg% purprogalin ether solution at 420 nm.

The electrochemical measurement kit of the present invention contains an aminoanthraquinone-based compound of the present invention or an electrode modifier containing the aminoanthraquinone-based compound in an amount sufficient for at least one assay. Typically, the electrochemical measurement kits of the invention, in addition to the aminoanthraquinone compounds of the invention, are oxidoreductases, buffers required for the assay, substrate standard solutions for making calibration curves, and Includes guidelines. For example, the oxidoreductase may be GDH, in which case the substrate standard solution can be a glucose standard solution.

In certain embodiments, the electrochemical measurement kit of the present invention comprises the aminoanthraquinone compound of the present invention and an oxidoreductase as the same reagent. In another embodiment, the electrochemical measurement kit of the present invention contains an aminoanthraquinone compound and an oxidoreductase as separate reagents. In another embodiment, the oxidoreductase may be immobilized on an electrode, and the electrochemical measurement kit of the present invention used for such an electrode contains an aminoanthraquinone-based compound as a single reagent. However, the term "single reagent" as used herein does not mean that the reagent does not contain a substance other than an aminoanthraquinone compound. The single reagent may include a suitable medium such that the aminoanthraquinone compounds of the present invention dissolve in the single reagent. The medium may be any medium as long as it dissolves the aminoanthraquinone compound of the present invention, and examples thereof include, but are not limited to, water, methanol, ethanol, propanol, acetone, and acetonitrile. The aminoanthraquinone compounds of the present invention can be used in various forms, for example, as a powder solid reagent, as a reagent immobilized on a bead or an electrode surface, or as a solution in a suitable storage solution, for example, as a light-shielded solution. Can be provided.

One example of an electrochemical measurement is the measurement of glucose concentration. In the case of colorimetric electrochemical measurement, the glucose concentration can be measured, for example, as follows. Glucose dehydrogenase (GDH) is used for the reaction layer for electrochemical measurement, and N- (2-acetamide) imide diacetic acid (ADA) and bis (2-hydroxyethyl) iminotris (hydroxymethyl) methane (Bis) are used as reaction promoters. A liquid or solid composition containing one or more substances selected from the group consisting of -Tris), sodium carbonate and imidazole is retained. Here, a pH buffer and a color-developing reagent (color-changing reagent) are added as needed. A sample containing glucose is added thereto, and the mixture is reacted for a certain period of time. During this period, the absorbance corresponding to the maximum absorption wavelength of the dye produced by polymerization or reduced dye by directly receiving electrons from GDH is monitored. Calibration prepared in advance using a glucose solution of standard concentration from the rate of change in absorbance per hour for the rate method and the change in absorbance up to the point when all glucose in the sample was oxidized for the endpoint method. The glucose concentration in the sample can be calculated based on the ion curve.

As a color-developing reagent (color-changing reagent) that can be used in this method, for example, 2,6-dichloroindophenol (DCIP) can be added as an electron acceptor, and glucose can be quantified by monitoring the decrease in absorbance at 600 nm. .. Further, it is possible to determine the amount of diformazan produced by adding nitrotetrazolium blue (NTB) as a color-developing reagent and measuring the absorbance at 570 nm, and to calculate the glucose concentration. Needless to say, the color-developing reagent (color-changing reagent) used is not limited to these.

In a certain embodiment, the property that the compound of the present invention is adsorbed on the electrode can be utilized to prepare a sensor capable of detecting the compound of the present invention. An electrode, for example, a carbon electrode, is inserted into a solution containing the compound of the present invention, for example, 1-acetamide-4-hydroxyanthraquinone, 1-amino-4-hydroxyanthraquinone, or 2-amino-3-hydroxyanthraquinone. Alternatively, the amount of the compound of the present invention can be quantitatively or qualitatively detected by electrochemical measurement, for example, CV or chronoamperometry, by inserting the compound into another measurement solution after a lapse of a certain period of time.

(Enzyme sensor)
In certain embodiments, the present invention provides an enzyme sensor comprising an electrode in which an oxidoreductase is immobilized and an electrode modifier of the present invention is adsorbed. Examples of the electrode of the enzyme sensor include a carbon electrode, a gold electrode, a platinum electrode, and the like, and an oxidoreductase can be applied or immobilized on the electrode. Furthermore, as conductive materials, Co, Pd, Rh, Ir, Ru, Os, Re, Ni, Cr, Fe, Mo, Ti, Al, Cu, V, Nb, Zr, Sn, In, Ga, Mg, Pb, Metal fine particles containing at least one element of Au, Pt, and Ag may be contained, and these may be alloys or plated. Examples of carbon include carbon nanotubes, carbon black, graphite, fullerenes, and derivatives thereof. For the method of immobilizing the oxidoreductase, refer to the paragraph "Immersion of enzyme or oxidoreductase" above.

In certain embodiments, an example of the enzyme sensor of the present invention is a glucose sensor. The glucose sensor may have the aminoanthraquinone compound of the invention adsorbed on the electrode and GDH or glucose oxidase (GOD) immobilized on the electrode. The glucose sensor can be used for continuous blood glucose measurement and continuous glucose monitoring.

The electrode, enzyme sensor, and composition for electrochemical measurement of the present invention can be used for various electrochemical measurement methods by using a potentiostat, galvanostat, or the like. Electrochemical measurement methods include various methods such as amperometry, such as chronoamperometry, potential step chronoamperometry, voltammetry, such as cyclic voltammetry, differential pulse voltammetry, potentialometry, and coulometry. For example, when the substrate to be measured is glucose, the glucose concentration in the sample can be calculated by measuring the current when glucose is reduced by the amperometry method. The applied voltage can be, for example, -1000 mV to +1000 mV (vs. Ag / AgCl), although it depends on the conditions and the setting of the device.

Whether or not the test compound is adsorbed on the electrode can be confirmed by cyclic voltammetry. Change the sweep rate, for example, in the range of 4 mV / sec to 200 mV / sec, and _{investigate how the maximum value of oxidation current (I Omax} ) changes. It is generally known that when the mediator is adsorbed on the electrode, the sweep rate of cyclic voltammetry and _{the value of I Omax} are in a proportional relationship. If the mediator is diffused, I _Omax is proportional to the sweep rate to the 0.5th power. From _{the relationship between I Omax} and the sweep rate, it is determined whether the compound is adsorbed or diffused.

An example of an electrochemical measurement is the electrochemical measurement of glucose. In certain embodiments, the present invention comprises contacting a sample that may contain glucose with an aminoanthraquinone compound with purified glucose oxidase or purified glucose dehydrogenase, and measuring current. Provided is a chemical measurement method. The aminoanthraquinone-based compound may be present in the solution or adsorbed on the electrode, and the enzyme may be immobilized on the electrode.

For example, the electrochemical measurement of glucose concentration can be performed as follows. A buffer solution is placed in a constant temperature cell and maintained at a constant temperature. A GDH or GOD-immobilized electrode is used as the working electrode, and a counter electrode (for example, a platinum electrode) and a reference electrode (for example, an Ag / AgCl electrode or an Ag / Ag + electrode) are used. The aminoanthraquinone compound of the present invention is added to the reaction solution. A constant voltage is applied to the carbon electrode, and after the current becomes steady, a sample containing glucose is added and the increase in current is measured. The glucose concentration in the sample can be calculated according to the calibration curve prepared with the glucose solution of the standard concentration. The applied potential can be, for example, +800 mV or less, + 700 mV or less, + 600 mV or less, + 500 mV or less, + 400 mV or less, + 300 mV or less, + 200 mV or less, + 100 mV or less, + 50 mV or less, and -200 mV or more. , -100 mV or more, -50 mV or more, for example 0 mV or more, for example + 800 mV to -200 mV, + 800 mV to -100 mV, + 800 mV to -50 mV, + 600 mV to 0 mV, + 500 mV to 0 mV, + 400 mV. It can be ~ 0 mV, +300 mV ~ 0 mV, +200 mV ~ 0 mV (all with respect to the silver chloride reference electrode). The pH of the measurement solution containing glucose can be in the pH range of 3-10. For example, the values are pH5, pH6, pH7, pH8, pH9, and pH10, and the solution may contain glycine, acetic acid, citric acid, phosphoric acid, carbonic acid, a good buffer, and the like as buffers. Even when an enzyme other than GDH or GOD and a substrate other than glucose are used, the pH can be appropriately changed for measurement.

As a specific example, an aminoanthraquinone compound such as 1-acetamide-4-hydroxyanthraquinone, 1-amino-4-hydroxyanthraquinone, or 2-amino-3-hydroxyanthraquinone is previously immobilized on a glassy carbon (GC) electrode. Then, 0.2U to 150U, for example 0.5U to 100U of GDH or GOD is immobilized, and the response current value with respect to the glucose concentration is measured. Add 10.0 ml of 100 mM potassium phosphate buffer (pH 6.0) into the electrolytic cell. Connect the GC electrode to the potentiostat BAS100B / W (manufactured by BAS), stir the solution at 37 ° C. and apply + 600 mV to the silver chloride reference electrode. Add 1M D-glucose solution to these systems to a final concentration of 5, 10, 20, 30, 40, 50 mM, and measure the steady-state current value for each addition. This current value is plotted against a known glucose concentration (5, 10, 20, 30, 40, 50 mM) to create a calibration curve. This makes it possible to quantify glucose with a GDH or GOD enzyme-immobilized electrode.

Another example of electrochemical measurement is the electrochemical measurement of hydrogen peroxide. In certain embodiments, the present invention comprises a step of contacting a sample that may contain hydrogen peroxide with an aminoanthraquinone compound and a purified peroxidase, and a step of measuring an electric current. I will provide a. The aminoanthraquinone-based compound may be present in the solution or adsorbed on the electrode, and the enzyme may be immobilized on the electrode.

For example, the electrochemical measurement of the hydrogen peroxide concentration can be performed as follows. A buffer solution is placed in a constant temperature cell and maintained at a constant temperature. An electrode on which peroxidase is immobilized is used as a working electrode, and a counter electrode (for example, a platinum electrode) and a reference electrode (for example, an Ag / AgCl electrode or an Ag / Ag + electrode) are used. The aminoanthraquinone compound of the present invention is added to the reaction solution. A constant voltage is applied to the carbon electrode, and after the current becomes steady, a sample containing hydrogen peroxide is added and the decrease in current is measured. The hydrogen peroxide concentration in the sample can be calculated according to the calibration curve prepared with the standard concentration hydrogen peroxide solution. The applied potentials are, for example, + 600 mV or less, + 500 mV or less, + 400 mV or less, + 300 mV or less, + 200 mV or less, + 100 mV or less, 0 mV or less, -100 mV or less, -200 mV or less, -300 mV or less, -400 mV or less,- It can be 500 mV or less, -1000 mV or more, -900 mV or more, for example -800 mV or more, for example + 600 mV to -800 mV, + 600 mV to -700 mV, + 500 mV to -600 mV, + 400 mV to- It can be 500 mV, +300 mV to -400 mV, +200 mV to -300 mV, + 100 mV to -200 mV, + 100 mV to -100 mV (all with respect to the silver chloride reference electrode). The pH of the measurement solution containing hydrogen peroxide can be in the range of pH 3-10. For example, the values are pH5, pH6, pH7, pH8, pH9, and pH10, and the solution may contain glycine, acetic acid, citric acid, phosphoric acid, carbonic acid, a good buffer, and the like as buffers. Even when an enzyme other than peroxidase and a substrate other than hydrogen peroxide are used, the pH can be appropriately changed for measurement.

As a specific example, an aminoanthraquinone compound such as 1-acetamide-4-hydroxyanthraquinone, 1-amino-4-hydroxyanthraquinone, or 2-amino-3-hydroxyanthraquinone is previously immobilized on a glassy carbon (GC) electrode. Then, 0.2U to 150U, for example 0.5U to 100U of peroxidase is immobilized, and the response current value with respect to the glucose concentration is measured. Add 10.0 ml of 100 mM potassium phosphate buffer (pH 7.0) into the electrolytic cell. Connect the GC electrode to the potentiostat BAS100B / W (manufactured by BAS) and apply + 600 mV to the silver chloride reference electrode. Add hydrogen peroxide solution to these systems so that the final concentration is 0.05, 0.1, 0.2, 0.5, 1 mM, and measure the steady-state current value for each addition. This current value is plotted against known hydrogen peroxide concentrations (0.05, 0.1, 0.2, 0.5, 1 mM) to create a calibration curve. This makes it possible to quantify hydrogen peroxide with the peroxidase enzyme-immobilized electrode.

Further, a printed electrode can be used for electrochemical measurement. This makes it possible to reduce the amount of solution required for measurement. Electrodes can be formed on an insulating substrate. Specifically, electrodes can be formed on a substrate by a photolithography technique or a printing technique such as screen printing, gravure printing, or flexographic printing. Examples of the material of the insulating substrate include silicon, glass, ceramic, polyvinyl chloride, polyethylene, polypropylene, polyester and the like, and those having strong resistance to various solvents and chemicals can be used. The area of the working electrode can be set according to the desired response current. For example, in one embodiment, the area of the working electrode is 1 mm ² or more, 1.5 mm ² or more, 2 mm ² or more, 2.5 mm ² or more, 3 mm ² or more, 4 mm ² or more, 5 mm ² or more, 6 mm ² or more, 7 mm ² or more, 8 mm. ² or more, 9 mm ² or more, 10 mm ² or more, 12 mm ² or more, 15 mm ² or more, 20 mm ² or more, 30 mm ² or more, 40 mm ² or more, 50 mm ² or more, 1 cm ² or more, 2 cm ² or more, 3 cm ² or more, 4 cm ² or more , 5 cm ² or more, for example 10 cm ² or more. In some embodiments, the area of the working electrode can be 10 cm ² or less, 5 cm ² or less, for example 1 cm ² or less. The opposite pole can be similar. In addition, carbon nanotubes, graphene, Ketjen black, etc. can be immobilized on the working surface to increase the apparent surface area. In this case, the apparent area can be increased 10 times or more, 50 times or more, 100 times or more, 1000 times or more.

In certain embodiments, the electrode modifiers, electrode adsorbents or electron transfer promoters of the present invention, when used with electrodes, have 0.1 pmol or more, 0.2 pmol or more, 0.3 pmol or more, per ^{1 cm 2 of working electrode area.} 0.4 pmol or more, 0.5 pmol or more, 1 pmol or more, 10 mmol or less, 5 mmol or less, 1 mmol or less, 800 μmol or less, 600 μmol or less, 500 μmol or less, 400 μmol or less, 300 μmol or less, 200 μmol or less, 100 μmol or less, 50 μmol or less, for example 0.1 pmol to 10 mmol, 0.1 pmol to 5 mmol, 0.2 pmol to 1 mmol, 0.3 pmol to 800 μmol, 0.4 pmol to 600 μmol, 0.5 pmol to 500 μmol, 0.6 pmol to 400 μmol, 0.7 pmol to 300 μmol, 0.8 pmol to 200 μmol, 0.9 pmol to 100 μmol, 1 pmol to 50 μmol Can be used in quantity. These values are for the case where the area of the working electrode is 1 cm ^2, and the apparent surface area increases when the area of the working electrode is increased or decreased, or when carbon nanotubes, graphene, etc. with a large specific surface area are used. However, the electrode modifier of the present invention with a corresponding amount of substance can be used. In certain embodiments, the electrode modifier, electrode adsorbent or electron transfer promoter of the present invention is 0.1 nmmol / L or more, 1 nmmol / L or more, 10 nmmol / L or more, 100 nmmol when used by being dissolved in an electrolyte. / L or more, 1 μmmol / L or more, 10 μmmol / L or more, 100 μmmol / L or more, 1 mmol / L or more, 10 mmol / L or more, 100 mmol / L or more, 1 mol / L or more, 10 mol / L or more, saturation concentration or less, 100 mol / It can be used at a concentration of L or less, for example, 1 mmol to saturation concentration and 10 mmol to 100 mol / L.

(Battery of the present invention)
In certain embodiments, the present invention provides a battery comprising the aminoanthraquinone compound of the present invention, an electrode modifier, an electrode adsorbent, or an electron transfer promoter. In certain embodiments, in the battery of the invention, the compounds of the invention are immobilized on the electrodes of the battery. In certain embodiments, the batteries of the present invention have an oxidoreductase. In certain embodiments, the redox enzyme may be immobilized on the electrodes of the battery. In certain embodiments, the present invention provides a power generation method using the batteries of the present invention.

(Battery of the present invention)
In certain embodiments, the present invention provides an anode or cathode for a fuel cell having the aminoanthraquinone-based compound of the present invention, an electrode modifier, an electrode adsorbent, or an electron transfer promoter, and a fuel cell having the anode or cathode. To do. In a certain embodiment, the present invention uses an aminoanthraquinone-based compound of the present invention, a power generation method using an electrode on which an aminoanthraquinone-based compound is adsorbed, or an oxidoreductase such as GDH or GOD is immobilized on an anode electrode to form an oxidoreductase. Provided is a power generation method using a corresponding substrate, for example, glucose as a fuel. When the above-mentioned oxidoreductase is immobilized, a compound serving as a substrate for the immobilized oxidoreductase can be used as a fuel. In certain embodiments, the present invention provides a flow battery comprising the aminoanthraquinone compound of the present invention, an electrode modifier, an electrode adsorbent, or an electron transfer promoter. The electrolyte solution contained in the electrolyte tank can be appropriately selected according to the battery, and may be an aqueous solution or an organic solvent. The pH of the electrolyte solution can be appropriately set in the range of pH 1 to 14.

In certain embodiments, the fuel cell of the present invention comprises an anode or cathode adsorbed by the aminoanthraquinone compound of the present invention, a fuel tank, a cathode, an anode having an oxidoreductase, and an electrolyte. Further, the fuel cell of the present invention can be provided with a load resistor between the anode and the cathode, if necessary, and can be provided with wiring for that purpose. In certain embodiments, the load resistance is part of the fuel cell of the present invention. In certain embodiments, the load resistance is not part of the fuel cell of the invention, and the fuel cell of the invention is configured to be connectable to a suitable load resistance. In the fuel cell of the present invention, the oxidoreductase constitutes a part of the anode. For example, the redox enzyme may be in close proximity to or in contact with the anode, may be immobilized, or may be adsorbed. The fuel tank contains a compound that serves as a substrate for oxidoreductase immobilized on an electrode. For example, if glucose dehydrogenase is immobilized on an electrode, the fuel can be glucose. In certain embodiments, the fuel cell of the present invention may have an ion exchange membrane that separates the anode and cathode. The ion exchange membrane can have pores of 1 nm to 20 nm. The anode can be a common electrode such as a carbon electrode. For example, electrodes made of conductive carbonaceous materials such as carbon black, graphite, activated carbon, carbon nanotubes, and graphene, and electrodes made of metals such as gold and platinum can be used. Specific examples thereof include carbon paper, carbon cloth, carbon felt, glassy carbon, and HOPG (highly oriented pyrolytic graphite). As the paired cathode, for example, an electrode catalyst generally used in a fuel cell such as platinum or a platinum alloy is used as a carbon such as carbon black, graphite, carbon cloth, carbon felt, activated carbon, carbon nanotube, and graphene. An electrode supported on a quality material or a conductor made of gold, platinum, etc., or a conductor made of an electrode catalyst itself such as platinum or a platinum alloy is used as a cathode electrode, and an oxidizing agent (cathode side substrate, oxygen, etc.) is used as an electrode. It can be in the form of supplying to the catalyst. In certain embodiments, the mercury electrode is removed from the electrodes of the present invention.

In another embodiment, the substrate reducing enzyme electrode can be used as the cathode paired with the anode composed of the substrate oxidizing enzyme electrode as described above. Examples of the redox enzyme that reduces the oxidizing agent include known enzymes such as peroxidase, laccase, and bilirubin oxidase. When an oxidoreductase is used as a catalyst for reducing the oxidant, a known electron transfer mediator may be used, if necessary. Examples of the oxidizing agent include oxygen and hydrogen peroxide.

In certain embodiments, an oxygen-selective membrane (eg, a dimethylpolysiloxane membrane) is placed around the cathode electrode to avoid the effects of impurities (such as ascorbic acid and uric acid) that interfere with the electrode reaction at the cathode. Can be done.

The power generation method of the present invention includes a step of supplying a compound serving as a substrate of the oxidoreductase as a fuel to the anode having the oxidoreductase. When fuel is supplied to the anode having oxidoreductase, the substrate is oxidized and the electrons generated at the same time are generated by the oxidoreductase, which is an electron transfer mediator that mediates electron transfer between the oxidoreductase and the electrode, for example. It is transferred to an aminoanthraquinone-based compound, and electrons are transferred to a conductive substrate (anode electrode) by the electron transfer mediator. An electric current is generated when electrons reach the cathode electrode from the anode electrode through wiring (external circuit). In certain embodiments, the power generation method of the present invention comprises a step of causing a redox reaction of an aminoanthraquinone compound. Since the aminoanthraquinone-based compound undergoes bond elimination with a proton by the transfer of electrons, it can be charged and discharged satisfactorily by a redox reaction.

^{The protons (H +} ) generated in the above process move to the cathode electrode in the electrolyte solution. Then, at the cathode electrode, the protons that have moved from the anode in the electrolyte solution, the electrons that have moved from the anode side through the external circuit, and the oxidizing agent (cathode side substrate) such as oxygen and hydrogen peroxide react. Water is produced. This can be used to generate electricity. An electrode containing an aminoanthraquinone-based compound may be arranged on the cathode side, and the oxidant of the aminoanthraquinone-based compound receives an electron and reacts with the reducing body to generate power.

(Organic Battery of the Present Invention)
In certain embodiments, the present invention provides an organic battery comprising the aminoanthraquinone compound of the present invention, an electrode modifier, an electrode adsorbent, or an electron transfer promoter. The electrode modifier, the electrode adsorbent, or the electron transfer promoter may be adsorbed on the electrode. Examples of the electrode material used for the organic battery include, but are not limited to, quinone, an indigo derivative, a benzoquinone compound having a methoxy group, indigocarmine, pentacentetron and the like. As the electrode material used for the organic radical battery, a compound having a nitroxy radical, for example, 2,2,6,6-tetramethylpiperidin-N-oxyl, which is bonded to a polymer, for example, polymethacrylate or acrylate, or lithium. However, it is not limited to this. See, for example, Polymer, Vol. 54, December, 2005, p.886. In an organic battery, the electrode modifier of the present invention can be used as an active material on the anode side and the cathode side.

The electrode modifier containing the aminoanthraquinone compound of the present invention and the electrode on which the electrode modifier is adsorbed can be used for various electrochemical measurements. Further, the electrode can be used as an enzyme sensor by immobilizing an oxidoreductase. Further, the electrode modifier containing the aminoanthraquinone compound of the present invention and the electrode to which the electrode modifier is adsorbed can be used for a fuel cell or an organic battery. These are examples, and the use of the electrode modifier containing the aminoanthraquinone compound of the present invention or the electrode on which the electrode modifier is adsorbed is not limited to this.

The aminoanthraquinone-based compound of the present invention may be adsorbed on the electrode as it is, or may be linked to the electrode by chemical modification with a cross-linking reagent or the like. Examples of the cross-linking reagent include, but are not limited to, 1-pyrenebutyric acid N-hydroxysuccinimide ester. Further, the aminoanthraquinone-based compound can be directly modified to the electrode by graft polymerization.

The present invention is further illustrated by the following examples. However, the technical scope of the present invention is not limited to those examples.

Materials and Methods Unless otherwise noted, materials and reagents are commercially available or obtained or prepared according to methods commonly used in the art and procedures in publicly known literature. As the single-walled carbon nanotubes, those manufactured by Sigma, Meijo Nanocarbon, or Zeon Nanotechnology were used. As the multi-walled carbon nanotubes, those manufactured by Sigma, Meijo Nanocarbon, or Kanto Chemical Co., Ltd. were used. The carbon nanotubes used were appropriately dispersed by sonication in an aqueous solution containing a low-molecular-weight surfactant, a water-soluble polymer, a water-soluble polysaccharide, and the like. Triton X-100, sodium dodecyl sulfate, etc. are used as the surfactant, but the surfactant is not limited to this. Compounds 1-acetamido-4-hydroxyanthraquinone, 1-amino-4-hydroxyanthraquinone, and 2-amino-3-hydroxyanthraquinone were commercially available from Tokyo Chemical Industry Co., Ltd.

Example 1. Confirmation of adsorptivity between aminoanthraquinone compounds and carbon electrodes Three types of aminoanthraquinone compounds (1-acetamide-4-hydroxyanthraquinone, manufactured by Tokyo Chemical Industry Co., Ltd., product code A1130), 1-amino-4-hydroxyanthraquinone, Tokyo Cyclic voltammetry (CV) was performed using printed electrodes using product code A0314) manufactured by Kasei Kogyo Co., Ltd., and 2-amino-3-hydroxyanthraquinone, product code A0315) manufactured by Tokyo Kasei Kogyo Co., Ltd. Specifically, a carbon working electrode ( ^{12.6 mm 2} ) and a silver reference electrode are printed on SCREEN-PRINTED ELECTRODES (DropSens, product number DRP-C110) with a final concentration of 10 μg / ml. -Add 15 μl of a 10% aqueous ethanol solution of acetamide-4-hydroxyanthraquinone, 1-amino-4-hydroxyanthraquinone, or 2-amino-3-hydroxyanthraquinone and 45 μl of 20 mM potassium phosphate buffer containing 1.5 M potassium chloride. did. After that, the electrode was connected to the ALS electrochemical analyzer 814D (BAS) using a dedicated connector (DRP-CAC, DropSens). Cyclic voltammetry was performed by sweeping the potential in a predetermined range using the working electrode, the reference electrode, and the counter electrode printed on the electrode. The range of potentials to be swept is + 300 mV to + 1000 mV (vs. Ag / AgCl) for 1-acetamido-4-hydroxyanthraquinone and + 200 mV to +800 mV (vs. Ag / AgCl) for 1-amino-4-hydroxyanthraquinone. , 2-Amino-3-hydroxyanthraquinone was adjusted from +200 mV to +600 mV (vs. Ag / AgCl). The sweep rate was changed in the range of 4 mV / sec to 200 mV / sec, and _{how the maximum value of oxidation current (I Omax} ) changed was investigated. It is generally known that when the mediator is adsorbed on the electrode, the sweep rate of cyclic voltammetry and _{the value of I Omax} are in a proportional relationship. If the mediator is diffused, I _Omax is proportional to the sweep rate to the 0.5th power.

Cyclic voltammetry using 1-acetamide-4-hydroxyanthraquinone was performed, and the results of plotting the _{sweep rate and I Omax are shown in FIG.} The value of R ^{2 was 0.9952.} On the other hand, when the results were plotted with the horizontal axis as the 0.5th power of the sweep rate, ^{the value of R 2} decreased to 0.9765. That is, it was confirmed that the sweep rate and I _Omax were in a proportional relationship, and it was shown that 1-acetamido-4-hydroxyanthraquinone was adsorbed on the carbon electrode.

FIG. 2 shows the results of a similar test using 1-amino-4-hydroxyanthraquinone instead of 1-acetamido-4-hydroxyanthraquinone. The value of R ^{2 was 0.9954.} On the other hand, when the results were plotted with the horizontal axis as the 0.5th power of the sweep rate, ^{the value of R 2} decreased to 0.9681. That is, it was confirmed that the sweep rate and IOmax were in a proportional relationship, and it was shown that 1-amino-4-hydroxyanthraquinone was adsorbed on the carbon electrode.

The results of conducting a similar test using 2-amino-3-hydroxyanthraquinone are shown in FIG. The value of R ^{2 was 0.9987.} On the other hand, when the results were plotted with the horizontal axis as the 0.5th power of the sweep rate, ^{the value of R 2} decreased to 0.9687. That is, it was confirmed that the sweep rate and IOmax were in a proportional relationship, and it was shown that 2-amino-3-hydroxyanthraquinone was adsorbed on the carbon electrode.

Comparative example (p-phenylenediamine)
10 μl of p-phenylenediamine 10% ethanol solution with a final concentration of 100 μg / ml and 10 μl of 100 mM potassium phosphate buffer (pH 7.0) were added dropwise to the above printed electrode, and the solution was -200 mV to + 400 mV (vs. Ag / Ag +). The result of cyclic voltammetry in which the voltage is swept in the range is shown in FIG. The value of R ^{2 was 0.9565.} On the other hand, when plotting the horizontal axis as the 0.5th power of the sweep speed, ^{the value of R 2} increased to 0.9899, _confirming that I Omax is proportional to the 0.5th power of the sweep speed. This indicates that p-phenylenediamine is not adsorbed on the electrode but is diffused.

Example 2. Electrochemical evaluation using aminoanthraquinone compounds
Cyclic voltammetry using a printed electrode was performed using FAD-dependent GDH and one of the aminoanthraquinone compounds of the present invention (1-amino-4-hydroxyanthraquinone). Specifically, a round carbon electrode (manufactured by Biodevice Technology Co., Ltd., DEP-Chip EP-PP) with a carbon working electrode (2.64 mm ² ) and a silver chloride reference electrode printed on it, with a dedicated connector. 50 mM phosphate containing 2 μl of 2000 U / ml FADGDH-AA (Kikkoman Biochemifa, product number 60100) solution, 1.5 M potassium chloride, connected to ALS electrochemical analyzer 814D (BAS). 10 μl of a 10% ethanol aqueous solution of 1-amino-4-hydroxyanthraquinone prepared in 8 μl of potassium buffer (pH 7.0) and 10 μg / ml was added dropwise onto the electrode. Then, cyclic voltammetry was performed in which the voltage was swept in the range of -400 mV to 800 mV (vs. Ag / AgCl). The sweep rate was 50 mV / sec. Subsequently, 2 μl of a 500 mM glucose aqueous solution was added, and cyclic voltammetry was performed in the same manner.

The oxidation current when + 600 mV was applied was 123 nA when glucose was not added, whereas it was 716 nA when glucose was added. Observation of the response current to glucose indicated that 1-amino-4-hydroxyanthraquinone was functioning as a mediator.

Subsequently, peroxidase (manufactured by Toyobo Co., Ltd., product number PEO-301, derived from Western wasabi) and three aminoanthraquinone compounds (1-acetamido-4-hydroxyanthraquinone, 1-amino-4-hydroxyanthraquinone, 2-amino- Cyclic voltammetry using a printed electrode was performed using 3-hydroxyanthraquinone). Specifically, 5 μl of 1100 U / ml peroxidase solution, 25 μl of 20 mM potassium phosphate buffer (pH 7.0) containing 1.5 M potassium chloride, and 15 μl of 10% ethanol solution of 10 μg / ml aminoanthraquinone compound. The solution was dropped onto the electrode, and cyclic voltammetry was performed with a sweep rate of 30 mV / sec and a sweep range of +200 mV to +800 mV (vs. Ag / AgCl). Furthermore, 5 μl of a 3 mM hydrogen peroxide aqueous solution was added, and cyclic voltammetry was carried out in the same manner. As a control experiment, the same test was carried out under the condition that the aminoanthraquinone compound was not contained, and the current values were compared. A wild-type enzyme may be used as the peroxidase.

The difference in reduction current between the presence and absence of hydrogen peroxide at +400 mV was 152 nA under the condition without aminoanthraquinone compounds, whereas it was 152 nA in the presence of 1-acetamide-4-hydroxyanthraquinone. It was 266nA, 284nA in the presence of 1-amino-4-hydroxyanthraquinone, and 241nA in the presence of 2-amino-3-hydroxyanthraquinone. In the presence of the aminoanthraquinone compound, the response current to hydrogen peroxide was increased as compared with the absence of the aminoanthraquinone compound, indicating that these compounds also function as mediators for peroxidase. Therefore, it was shown that the mediator of the present invention can be used not only for the anode electrode but also for the cathode electrode.

Example 3. Electrochemical evaluation of electrodes chemically modified with aminoanthraquinone compounds
80 μl of a multi-walled carbon nanotube solution (1.4 wt%, MWCNT solution) was applied to a 5 mm square carbon cloth (manufactured by Toyo Corporation) in multiple times. After it was sufficiently dried, it was washed with ultrapure water. Subsequently, 100 μg of 1-pyrenebutyrate N-hydroxysuccinimide was applied and sufficiently dried. Further, 20 μg of 1-amino-4-hydroxyanthraquinone, which is an aminoanthraquinone compound of the present invention dissolved in ethanol, was applied and dried, and then 20 μl of 20 mg / ml peroxidase was applied and dried at room temperature. The prepared electrode was washed with ultrapure water. Cyclic voltammetry was performed by immersing the three electrodes in 10 ml of 100 mM potassium phosphate buffer (pH 7) with the washed electrode as the working electrode, silver chloride as the reference electrode, and platinum as the counter electrode. The sweep speed was 20 mV / sec, and the sweep range was + 0 mV to + 700 mV (vs. Ag / AgCl). A hydrogen peroxide solution was added dropwise to a final concentration of 1 mM, and the difference in reduction current before and after the addition of hydrogen peroxide was observed. As a result, at + 0.3V, a reduction current that ^{was 0.54mA / cm 2 larger when hydrogen peroxide was added flowed than when hydrogen peroxide was not added.}

On the other hand, 20 μl of a multi-walled carbon nanotube dispersion solution is applied to a 0.5 mm square carbon cloth, dried at 60 ° C. for 1 hour or more, and 20 μl of a final concentration of 5 mg / ml peroxidase and 5% glutaraldehyde solution is applied. When the electrodes were prepared by drying at room temperature for a period of time, as a result of comparing the cyclic voltammograms before and after the addition of the hydrogen peroxide solution, the difference in reduction current was in the range of +200 mV to +700 mV (vs. Ag / AgCl). It was not seen.

Example 4. Construction of fuel cell
80 μl of a multi-walled carbon nanotube solution was applied to a 5 mm × 5 mm carbon cloth (manufactured by Toyo Corporation) in several batches, and dried at 60 ° C. After washing with pure water, the mixture was further dried, and the 1-amino-4-hydroxyanthraquinone of the present invention was adsorbed and fixed, and washed with ultrapure water. Subsequently, 20 μl of 20 mg / ml peroxidase (manufactured by Toyobo Co., Ltd., product number PEO-301) was applied, dried at 25 ° C., and the above carbon cloth was exposed to 25% glutaraldehyde vapor for 20 minutes to crosslink the peroxidase. It was fixed and used as a cathode electrode. Further, the multi-walled carbon nanotubes were applied to the carbon cloth and dried in the same procedure. As a mediator, after adsorbing and fixing N-isopropyl-N'-phenyl-p-phenylenediamine, 20 μl of 20 mg / ml FAD-dependent glucose dehydrogenase (GLD1, manufactured by Funakoshi) was applied, dried at 25 ° C, and 25. % Glutaraldehyde The anode electrode is obtained by cross-linking and fixing carbon cloth by exposing it to steam for 20 minutes. The above cathode electrode and anode electrode were immersed in 100 mM potassium phosphate buffer (pH 7) containing 200 mM D-glucose, and connected to a variable resistor and a potentiostat. The voltage was measured at the open circuit potential. As a result, the open circuit voltage was 0.37V. That is, it was shown that 1-amino-4-hydroxyanthraquinone functions as an active material for the cathode electrode. Further, when the above-mentioned cathode electrode and anode electrode were immersed in a 100 mM potassium phosphate buffer solution (pH 7) containing 200 mM D-glucose and 1 mM hydrogen peroxide, the open circuit voltage was 0.5 V. , 0.15mA / cm ² current flowed when connected to 10kΩ. Therefore, it was possible to construct a fuel cell using the electrode on which 1-amino-4-hydroxyanthraquinone of the present invention was adsorbed and fixed.

Further, single-walled carbon nanotubes can be used instead of the multi-walled carbon nanotubes.

By using the electrode modifier containing the aminoanthraquinone compound of the present invention or the electrode to which the electrode modifier is adsorbed, various electrochemical measurements of a battery or the like can be performed.

All publications, patents and patent applications mentioned or cited herein are incorporated herein by reference in their entirety.

Claims (18)

The structure of formula I, which has the property of adsorbing to the electrode without being bonded to the polymer or polymerized,

[In the formula, R ¹ is hydrogen, C _1-6 alkyl, optionally aryl or -C (= O) -C _1-6 alkyl optionally substituted with sulfo].
An electrode modifier comprising a compound having. The structure of formula I, which has the property of adsorbing to the electrode without being bonded to the polymer or polymerized,

[In the formula, R ¹ is hydrogen, C _1-6 alkyl, optionally aryl or -C (= O) -C _1-6 alkyl optionally substituted with sulfo].
An electron transfer promoter, which comprises a compound having. A battery comprising the electrode modifier according to claim 1 or the electron transfer promoter according to claim 2. The battery according to claim 3, wherein the compound is immobilized on an electrode of the battery. The battery according to claim 3 or 4, which comprises an oxidoreductase. The battery according to claim 5, wherein the redox enzyme is immobilized on an electrode. A composition comprising the electrode modifier according to claim 1 or the electron transfer promoter according to claim 2. An electrode comprising the electrode modifier according to claim 1 or the electron transfer promoter according to claim 2. The electrode according to claim 8 having an enzyme, or the composition according to claim 7, which comprises an enzyme. The electrode or composition according to claim 9, wherein the enzyme is an oxidoreductase. The electrode according to claim 10, wherein the redox enzyme is immobilized. A sensor containing the electrode modifier according to claim 1 or the electron transfer promoter according to claim 2, or an enzyme sensor having an electrode according to claim 10 or 11. The compound has the following structure

Has 1-acetamido-4-hydroxyanthraquinone,
The following structure

1-Amino-4-hydroxyanthraquinone with, or the following structure

Has 2-amino-3-hydroxyanthraquinone,
Or its salt, anhydride or solvate,
The electrode modifier according to claim 1,
The electron transfer promoter according to claim 2.
The battery according to any one of claims 3 to 6.
The composition according to claim 7, 9 or 10.
The electrode according to any one of claims 8 to 11, or the enzyme sensor according to claim 12. A method for manufacturing a battery, which comprises a step of using the electrode modifying agent according to claim 1 or the electron transfer accelerator according to claim 2. The method according to claim 14, further comprising a step of bringing the electrode modifier or the electron transfer accelerator into contact with the electrode of the battery. A power generation method using the battery according to any one of claims 3 to 6 and 13. The composition according to any one of claims 7, 9, 10 and 13.
An electrochemical measurement method using the electrode according to any one of claims 8 to 11 and 13 or the sensor according to claim 12 or 13. The electrode modifier according to claim 1 or 13.
The electron transport chain according to claim 2 or 13, or the composition according to any one of claims 7, 9, 10 and 13.
A method of modifying or modifying an electrode, which comprises the step of bringing the electrode into contact with the electrode.

Images