WO2005080953A1 - Urinary glucose biosensor - Google Patents

Urinary glucose biosensor Download PDF

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Publication number
WO2005080953A1
WO2005080953A1 PCT/JP2005/002391 JP2005002391W WO2005080953A1 WO 2005080953 A1 WO2005080953 A1 WO 2005080953A1 JP 2005002391 W JP2005002391 W JP 2005002391W WO 2005080953 A1 WO2005080953 A1 WO 2005080953A1
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Prior art keywords
glucose
urine
mediator
meter
biosensor according
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PCT/JP2005/002391
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French (fr)
Japanese (ja)
Inventor
Masao Gotoh
Hideaki Nakamura
Satoshi Koide
Fumiyo Kurusu
Isao Karube
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National Institute Of Advanced Industrial Science And Technology
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Priority to JP2006510213A priority Critical patent/JP4247844B2/en
Publication of WO2005080953A1 publication Critical patent/WO2005080953A1/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/001—Enzyme electrodes
    • C12Q1/005—Enzyme electrodes involving specific analytes or enzymes
    • C12Q1/006—Enzyme electrodes involving specific analytes or enzymes for glucose
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/483—Physical analysis of biological material
    • G01N33/487—Physical analysis of biological material of liquid biological material
    • G01N33/493—Physical analysis of biological material of liquid biological material urine

Definitions

  • the present invention relates to a urine glucose biosensor. More specifically, the present invention relates to a urine glucose biosensor for measuring glucose concentration in urine.
  • Normal urine also contains trace amounts of glucose, and basic data on urine is based on an average urine pH of 6.3 (4.8-7.5), urine sugar of 40-85 mgZ days, and urine volume of 600-1600 mlZ days. , The normal value of urinary sugar concentration is 20mg / dl.
  • urine glucose level 100-150mg / dl is a measure of lifestyle improvement. Consultation with a physician is required. Diabetes is the leading cause of blindness, amputation of limbs, neuropathy, kidney disease, etc.If urinary glucose can be easily determined, it will be easier to prevent those serious diseases, and the clinical significance of urinary glucose measurement Is in this respect.
  • a biosensor that integrates a membrane on which glucose oxidase (GOD) is immobilized and an electrode, oxidizes generated hydrogen peroxide with the electrode, and measures the current value is used. is there.
  • GOD glucose oxidase
  • the conventional urine glucose biosensor has a structure in which an electrode and a membrane on which an enzyme is immobilized are integrated, and is capable of measuring urine sugar many times. After being used in a solution, it is dried until the next use. In this state, the solution is repeatedly dried, and the state of the electrode, membrane and enzyme becomes unstable, and the reliability as a sensor is lost.Therefore, the electrode section and the enzyme-immobilized membrane are stable. In consideration of portability, it must always be stored in a state of being immersed in a buffer solution, and there is a problem in portability.
  • Patent document 1 JP-A-9-159645
  • An object of the present invention is to provide a urine glucose biosensor having excellent portability and having a function of measuring and displaying urine sugar. Means for solving the problem
  • An object of the present invention is to seal a carrier impregnated with glucose oxidase or glucose dehydrogenase and a mediator, or a glucose oxidase or glucose dehydrogenase and a mediator, in or near a pH electrode portion of a pH meter. This is achieved by a urine glucose biosensor provided with a reactant formed of a bag body.
  • the reagent in the carrier or bag is disposable each time, even if the reagent is stored in a dry state, the structural change of the enzyme is small, and the characteristics of the sensor become stable.
  • both the reactant and the pH measuring device are portable, it is possible to measure urinary sugar anytime and anywhere without attaching a buffer solution for preservation as with conventional measuring devices.
  • explanations and countermeasures for the measurement results are provided based on the concentration previously input to the pH meter or the data corresponding thereto, so that the user can know his / her daily health condition. It can be used for health management.
  • FIG. 1 is a graph showing the results obtained in Example 1.
  • FIG. 2 is a graph showing the results obtained in Example 2.
  • FIG. 3 is a graph showing the results obtained in Example 3.
  • FIG. 4 is a graph showing the results obtained in Example 4.
  • FIG. 5 is a graph showing the results obtained in Example 5.
  • mediator used together with glucose oxidase or glucose dehydrogenase
  • water-soluble ferricyanidani potassium, fuecsen, parabenzoquinone and the like are used as a mediator (electron carrier) used together with glucose oxidase or glucose dehydrogenase.
  • Reactants formed using these compounds are prepared by dissolving 13- 13250 units of glucose oxidase or glucose dehydrogenase and 3-280 mg of mediator in 0.1-10 ml of water.
  • the method is carried out by a method in which the reagent is added to the carrier and dried, and the reagent is impregnated in the carrier, or a method in which the reagent is packed in a bag and the reagent is sealed in the bag.
  • a desiccant and the like are appropriately packed.
  • the carrier include paper such as filter paper and non-woven cloth
  • examples of the bag include paper such as filter paper and non-woven cloth formed into a bag. Packs and soup packs can be used as they are.
  • the desiccant silica gel, activated alumina, zeolite, calcium chloride and the like are used.
  • the pH meter is not particularly limited as long as it can achieve the intended purpose when measuring the pH, but a portable one is preferably used.
  • a portable pH meter a commercially available product, for example, a simple pH meter Twin pH B-211 (glass electrode) manufactured by HORIBA, Ltd. is used as it is.
  • the pH electrode is preferably a glass electrode or an ion-sensitive field effect transistor (ISFET)!
  • Urine sugar concentration is measured, for example, when a pH electrode unit is provided at the bottom of a spoon-shaped container, a carrier impregnated with an enzyme and a mediator, or a bag containing the enzyme and a mediator is placed on the electrode. It is carried out by allowing the reactant to stand completely and urine completely on the reactant as described above, or by immersing the reactant in urine and measuring the pH after a certain period of time. Also, urine is collected in an appropriate container, and then a pH electrode is put into the urine, and the reactant is put there, and the pH after a certain time is determined.
  • Non-Patent Document 1 discloses a method using a pH electrode. It is shown that the enzyme is immobilized using a membrane on the surface of the However, in practice, the change in pH was so small that it could not be put to practical use.
  • Non-Patent Document 1 Anal.Chem., 57, 1920 (1985)
  • the present invention is to use a mediator to amplify the action of an enzyme with the mediator, increase the pH change, and make it possible to measure the pH with a sensitivity using a pH meter. That is, the dissolved oxygen that glucose oxidase normally uses as a type of mediator is
  • the concentration force in an aqueous solution is about ppm, and the force fluctuates.
  • the mediator is artificially supplied in a high concentration state so that it does not depend on the dissolved oxygen concentration.
  • the amount of dalconic acid generated is increased so that it can be detected with a pH electrode.
  • Glucose dehydrogenase does not depend on dissolved oxygen, but acts to increase the amount of dalconic acid generated by the action of the mediator.
  • pH which is the display value of the pH measuring device main body
  • a calibration curve indicating the relationship between pH and urine glucose concentration is stored in the pH measuring device main body, and measurement is displayed instead of urine glucose concentration. It can also be a device.
  • the pH of urine varies depending on individuals and the environment at the time of collection, by inserting a calibration curve corresponding to the initial pH in the main unit, specifically, the pH of urine can be measured first with the main unit of the pH meter. Then, the pH value after a certain period of time is detected and stored in the main body of the pH measuring instrument. The difference can be eliminated.
  • a symbol such as "Normal value” is displayed together with the urine glucose concentration in Z or alone.
  • aqueous solution was prepared by dissolving 1325 units of glucose oxidase (product of Sigma G-7016) and 28 mg of potassium ferricyanide (product of Wako Pure Chemical Industries) in 1 ml of pure water. Filter paper product qualitative filter paper No.1) was dropped and infiltrated, and dried in a refrigerator for a while to obtain a carrier.
  • Control urine Bio'Rad Laboratories product Quantitative Urine Control Glucose (Sigma product) was added to "Nomal" at 19.2, 28.6, 48.1, 53, 98 and 200.8 mg / dl to prepare urine standard samples with different glucose concentrations. It should be noted that the initial P H of urine standard sample of all and was 6.1.
  • Example 1 the reagents of glucose oxidase and potassium ferricyanide were used in a volume change of 1: 1. The obtained results are shown in Fig. 2, and it was confirmed that the measured values after 60 seconds showed calibration.
  • Example 1 the reagents of glucose oxidase and potassium ferricyanide were used in a state where the reagent volumes were changed to ⁇ times. The obtained results are shown in Fig. 3, and it was confirmed that the measured values after 30 seconds showed calibration.
  • Example 1 the reagents used for glucose oxidase and potassium ferricyanide were changed to twice the reagent capacity. The results obtained are shown in FIG. 4, and a clear difference between the concentrations appears after 60 seconds as a whole, indicating that the urinary glucose concentration can be measured.
  • the calibration range was approximately 50 to 200 mg Zdl.
  • Example 1 instead of the carrier, a product in which 3975 units of glucose oxidase (G-7016) and 84 mg of potassium potassium were used in a black tea pack (Lipton) was used. The obtained results are shown in FIG. 5, and a clear difference appeared for each concentration after 60 seconds as a whole, indicating that the urinary glucose concentration can be measured. Calibration range is about 30 to 200mgZdl.

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Abstract

A urinary glucose biosensor comprising a pH meter and, disposed on a portion of pH electrode or in the vicinity thereof, a reactant consisting of a bag having glucose oxidase or glucose dehydrogenase and a mediator sealed therein, or a carrier impregnated with glucose oxidase or glucose dehydrogenase and a mediator. In the use of the urinary glucose biosensor, the reagents of the carrier or bag are disposed of every time. Thus, even when the reagents are stored in the dry state, any structural change of the enzymes is slight, thereby stabilizing the performance of the sensor. Consequently, both the reactant and the pH meter are portable, so that as different from the conventional measuring instruments, it is not needed to add any buffer for storage and urinary glucose measurement can be performed at any time in any place.

Description

明 細 書  Specification
尿糖バイオセンサ  Urine sugar biosensor
技術分野  Technical field
[0001] 本発明は、尿糖バイオセンサに関する。さらに詳しくは、尿中のグルコース濃度を測 定する尿糖バイオセンサに関する。  The present invention relates to a urine glucose biosensor. More specifically, the present invention relates to a urine glucose biosensor for measuring glucose concentration in urine.
背景技術  Background art
[0002] 正常尿中にもグルコースが微量に存在し、尿の基本的データは、尿の pHが平均値 6.3(4.8— 7.5)、尿糖が 40— 85mgZ日、尿量が 600— 1600mlZ日、尿糖濃度正常値 カ^ー 20mg/dlである。糖質代謝異常によって血糖値が上昇した場合、上記尿糖濃度 も上がりそれは糖尿病とよばれ、一般に尿糖値力 100— 150mg/dlは生活習慣改善の 目安とされ、それ以上検出された場合は医師への相談が必要とされている。糖尿病 は、失明、手足の切断、神経障害、腎臓疾患等の原因の第一位であり、尿糖値を容 易に判別できればそれらの重大な疾患を防ぎやすくなり、尿糖測定の臨床的意義は この点にあるといえる。  [0002] Normal urine also contains trace amounts of glucose, and basic data on urine is based on an average urine pH of 6.3 (4.8-7.5), urine sugar of 40-85 mgZ days, and urine volume of 600-1600 mlZ days. , The normal value of urinary sugar concentration is 20mg / dl. When blood sugar level rises due to abnormal carbohydrate metabolism, the above-mentioned urinary glucose level also rises, which is called diabetes. Generally, urine glucose level of 100-150mg / dl is a measure of lifestyle improvement. Consultation with a physician is required. Diabetes is the leading cause of blindness, amputation of limbs, neuropathy, kidney disease, etc.If urinary glucose can be easily determined, it will be easier to prevent those serious diseases, and the clinical significance of urinary glucose measurement Is in this respect.
[0003] 従来の尿糖バイオセンサとしては、グルコースォキシダーゼ (GOD)を固定ィ匕した膜 と電極を一体とし、発生する過酸化水素を電極で酸化し、その電流値を測定するバ ィォセンサがある。  [0003] As a conventional urine glucose biosensor, a biosensor that integrates a membrane on which glucose oxidase (GOD) is immobilized and an electrode, oxidizes generated hydrogen peroxide with the electrode, and measures the current value is used. is there.
[0004] しかし、従来の尿糖バイオセンサは、電極と酵素を固定ィ匕した膜を一体とし、尿糖を 何回も測定できる構造であり、溶液中で使用した後、次の使用まで乾燥状態でおくと 、溶液 '乾燥の繰り返しになり、電極、膜、酵素の状態が不安定になって、センサとし ての信頼性を欠くことになることから、電極部と酵素固定化膜は安定性を考慮して、 常に緩衝液に浸した状態で保存しなければならず、携帯性に課題があった。  [0004] However, the conventional urine glucose biosensor has a structure in which an electrode and a membrane on which an enzyme is immobilized are integrated, and is capable of measuring urine sugar many times. After being used in a solution, it is dried until the next use. In this state, the solution is repeatedly dried, and the state of the electrode, membrane and enzyme becomes unstable, and the reliability as a sensor is lost.Therefore, the electrode section and the enzyme-immobilized membrane are stable. In consideration of portability, it must always be stored in a state of being immersed in a buffer solution, and there is a problem in portability.
特許文献 1：特開平 9— 159645号公報  Patent document 1: JP-A-9-159645
発明の開示  Disclosure of the invention
発明が解決しょうとする課題  Problems to be solved by the invention
[0005] 本発明の目的は、携帯性にすぐれ、尿糖の測定表示機能を有する尿糖バイオセン サを提供することにある。 課題を解決するための手段 An object of the present invention is to provide a urine glucose biosensor having excellent portability and having a function of measuring and displaying urine sugar. Means for solving the problem
[0006] 力かる本発明の目的は、 pH測定器の pH電極部またはその付近に、グルコースォキ シダーゼまたはグルコースデヒドロゲナーゼとメディエーターを含浸させた担体、ある いはグルコースォキシダーゼまたはグルコースデヒドロゲナーゼとメディエーターを封 入した袋体からなる反応体を設置した尿糖バイオセンサによって達成される。  [0006] An object of the present invention is to seal a carrier impregnated with glucose oxidase or glucose dehydrogenase and a mediator, or a glucose oxidase or glucose dehydrogenase and a mediator, in or near a pH electrode portion of a pH meter. This is achieved by a urine glucose biosensor provided with a reactant formed of a bag body.
発明の効果  The invention's effect
[0007] 担体または袋体中の試薬は 1回毎に使い捨てるため、試薬を乾燥状態で保存して も酵素の構造変化が少なぐセンサの特性も安定ィ匕するようになる。その結果、反応 体および pH測定器はいずれも携帯できることから、従来の測定機器のように、保存用 の緩衝液を付帯することなぐいつでもどこでも尿糖の測定が可能である。また、 pH 測定器に予め入力されていた濃度またはこれに対応したデータにより、結果の表示 の際、測定結果に対する説明や対処法が提供されるので、日常の健康状態を知るこ とができ、健康管理に役立てることができる。  [0007] Since the reagent in the carrier or bag is disposable each time, even if the reagent is stored in a dry state, the structural change of the enzyme is small, and the characteristics of the sensor become stable. As a result, since both the reactant and the pH measuring device are portable, it is possible to measure urinary sugar anytime and anywhere without attaching a buffer solution for preservation as with conventional measuring devices. Also, when the results are displayed, explanations and countermeasures for the measurement results are provided based on the concentration previously input to the pH meter or the data corresponding thereto, so that the user can know his / her daily health condition. It can be used for health management.
図面の簡単な説明  Brief Description of Drawings
[0008] [図 1]実施例 1で得られた結果を示すグラフである。 FIG. 1 is a graph showing the results obtained in Example 1.
[図 2]実施例 2で得られた結果を示すグラフである。  FIG. 2 is a graph showing the results obtained in Example 2.
[図 3]実施例 3で得られた結果を示すグラフである。  FIG. 3 is a graph showing the results obtained in Example 3.
[図 4]実施例 4で得られた結果を示すグラフである。  FIG. 4 is a graph showing the results obtained in Example 4.
[図 5]実施例 5で得られた結果を示すグラフである。  FIG. 5 is a graph showing the results obtained in Example 5.
発明を実施するための最良の形態  BEST MODE FOR CARRYING OUT THE INVENTION
[0009] グルコースォキシダーゼまたはグルコースデヒドロゲナーゼとともに用いられるメディ エーター (電子伝達体)としては、水溶性のフェリシアンィ匕カリウム、フエ口セン、パラべ ンゾキノンなどが用いられる。これらを用いた反応体の形成は、グルコースォキシダー ゼまたはグルコースデヒドロゲナーゼ 133— 13250単位およびメデイエ一ター 3— 280mgを水 0.1— 10mlに溶解したもの 10— 1000 μ 1を、デスペンサなどにより担体上に 滴下し、乾燥して、試薬を担体に含浸させる方法または試薬を袋体中にパックして、 試薬を袋体に封入する方法等によって行われる。このとき、試薬の安定性の観点力も 、好ましくは乾燥剤などが適宜パックされる。ここで、担体としては、ろ紙などの紙、不 織布等が挙げられ、袋体としては、ろ紙などの紙、不織布等を袋状としたもの等が挙 げられ、例えば市販されている紅茶用パック、だし用パック等をそのまま用いることが できる。また、乾燥剤としてはシリカゲル、活性アルミナ、ゼォライト、塩化カルシウム 等が用いられる。 [0009] As a mediator (electron carrier) used together with glucose oxidase or glucose dehydrogenase, water-soluble ferricyanidani potassium, fuecsen, parabenzoquinone and the like are used. Reactants formed using these compounds are prepared by dissolving 13- 13250 units of glucose oxidase or glucose dehydrogenase and 3-280 mg of mediator in 0.1-10 ml of water. The method is carried out by a method in which the reagent is added to the carrier and dried, and the reagent is impregnated in the carrier, or a method in which the reagent is packed in a bag and the reagent is sealed in the bag. At this time, the viewpoint of the stability of the reagent Preferably, a desiccant and the like are appropriately packed. Here, examples of the carrier include paper such as filter paper and non-woven cloth, and examples of the bag include paper such as filter paper and non-woven cloth formed into a bag. Packs and soup packs can be used as they are. As the desiccant, silica gel, activated alumina, zeolite, calcium chloride and the like are used.
[0010] この反応体を尿中に完全に浸漬または反応体の全体をおおうように尿がかけられる と、尿中の尿糖は、反応体中のグルコースォキシダーゼとフェリシアン化カリウムの作 用によりダルコン酸を発生させる。 jS .D-グルコース + 2Fe(CN)6 + H20 > [0010] When this reactant is completely immersed in urine or urine is applied so as to cover the whole of the reactant, urine sugar in the urine is converted to glucose by the action of glucose oxidase and potassium ferricyanide in the reactant. Generates acid. jS .D-glucose + 2Fe (CN) 6 + H 20 >
グノレコン酸 + 2Fe(CN)6—― + 2H+ このダルコン酸の発生を pH電極で検知することにより、間接的に尿糖濃度を測定す ることができる。グルコースデヒドロゲナーゼも、同様の反応機構でダルコン酸を発生 させる。 Gunoreconic acid + 2Fe (CN) 6 —— + 2H + By detecting the generation of this dalconic acid with a pH electrode, the urinary glucose concentration can be measured indirectly. Glucose dehydrogenase also generates dalconic acid by a similar reaction mechanism.
[0011] pH測定器は、 pHを測定するといつた目的を達成しうるものであれば特に限定されな いが、好ましくは携帯性のものが使用される。このような携帯型 pHメーターとしては、 市販品、例えば堀場製作所製品簡易 pHメーター Twin pH B-211 (ガラス電極)など がそのまま用いられる。 pH電極は、ガラス電極またはイオン感応性電界効果型トラン ジスター（ISFET)が好まし!/、。  [0011] The pH meter is not particularly limited as long as it can achieve the intended purpose when measuring the pH, but a portable one is preferably used. As such a portable pH meter, a commercially available product, for example, a simple pH meter Twin pH B-211 (glass electrode) manufactured by HORIBA, Ltd. is used as it is. The pH electrode is preferably a glass electrode or an ion-sensitive field effect transistor (ISFET)!
[0012] 尿糖濃度の測定は、例えば pH電極部がスプーン状の容器の底部に設置されてい る場合、酵素とメディエーターを含浸させた担体、もしくは酵素とメディエーターを含 む袋体を電極上に静置し、そこに上述した如く反応体に完全に尿をかけ、または反 応体を尿に浸漬し、一定時間後の pHを測定することにより行われる。また、適当な容 器に尿を採取し、次いで pH電極を尿中に入れ、そこに反応体を入れて一定時間後 の pHを柳』定することちでさる。  [0012] Urine sugar concentration is measured, for example, when a pH electrode unit is provided at the bottom of a spoon-shaped container, a carrier impregnated with an enzyme and a mediator, or a bag containing the enzyme and a mediator is placed on the electrode. It is carried out by allowing the reactant to stand completely and urine completely on the reactant as described above, or by immersing the reactant in urine and measuring the pH after a certain period of time. Also, urine is collected in an appropriate container, and then a pH electrode is put into the urine, and the reactant is put there, and the pH after a certain time is determined.
[0013] ここで、尿糖にグルコースォキシダーゼを作用させて生じるダルコン酸を pHにより測 定可能であることは理論的に示されており、非特許文献 1には、 pH電極を用いてそ の表面に膜を用いて酵素を固定ィ匕し、グルコースノィォセンサとすることが示されて いるものの、実際には pH変化が小さく実用化できるものではな力つた。 非特許文献 1 : Anal. Chem., 57, 1920 (1985) [0013] Here, it has been theoretically shown that dalconic acid generated by allowing glucose oxidase to act on urinary sugar can be measured by pH, and Non-Patent Document 1 discloses a method using a pH electrode. It is shown that the enzyme is immobilized using a membrane on the surface of the However, in practice, the change in pH was so small that it could not be put to practical use. Non-Patent Document 1: Anal.Chem., 57, 1920 (1985)
[0014] 本発明は、メディエーターを用いることにより、酵素の作用をメディエーターで増幅 し、 pH変化を大きくして、感度よく pH測定器で測定可能としたものである。すなわち、 通常グルコースォキシダーゼがー種のメディエーターとして使用している溶存酸素は[0014] The present invention is to use a mediator to amplify the action of an enzyme with the mediator, increase the pH change, and make it possible to measure the pH with a sensitivity using a pH meter. That is, the dissolved oxygen that glucose oxidase normally uses as a type of mediator is
、水溶液中での濃度力 ppm程度であり、し力も変動するものであるのに対し、本発明 においては人工的にメディエーターを高濃度の状態で供給することにより、溶存酸素 濃度に依存せず、ダルコン酸の発生量を多くして、 pH電極で検知できるようにしたも のである。グルコースデヒドロゲナーゼの場合には、溶存酸素に依存しないが、やは りメディエーターの作用により、ダルコン酸の発生量を多くする働きがある。 In contrast, the concentration force in an aqueous solution is about ppm, and the force fluctuates.On the other hand, in the present invention, the mediator is artificially supplied in a high concentration state so that it does not depend on the dissolved oxygen concentration. The amount of dalconic acid generated is increased so that it can be detected with a pH electrode. Glucose dehydrogenase does not depend on dissolved oxygen, but acts to increase the amount of dalconic acid generated by the action of the mediator.
[0015] ここで pH測定器本体の表示値である pHに代えて、 pH測定器本体に pHと尿糖濃度 の関係を示す検量線を記憶させておき、尿糖濃度にかえて表示する計測機器とする こともできる。また、尿の pHは個人差、採取時の環境差があるため、初期 pHに対応し た検量線を本体に入れておくことにより、具体的には尿の pHを最初に pH測定器本体 で検知し、次 、で一定時間後の pH値をあら力じめ pH測定器本体に記憶されて 、た 初期 pH毎の検量線に適用して尿糖濃度を表示することにより、このような個人差を解 消することができる。  [0015] Here, instead of pH, which is the display value of the pH measuring device main body, a calibration curve indicating the relationship between pH and urine glucose concentration is stored in the pH measuring device main body, and measurement is displayed instead of urine glucose concentration. It can also be a device. Also, since the pH of urine varies depending on individuals and the environment at the time of collection, by inserting a calibration curve corresponding to the initial pH in the main unit, specifically, the pH of urine can be measured first with the main unit of the pH meter. Then, the pH value after a certain period of time is detected and stored in the main body of the pH measuring instrument. The difference can be eliminated.
[0016] さらに表示内容としては、尿糖濃度の他に、尿糖濃度に対応した文章、単語、図形 [0016] Furthermore, in addition to the urine sugar concentration, sentences, words, and graphics corresponding to the urine sugar concentration are displayed.
、記号、例えば「正常値です」などのよう文章を尿糖濃度とともに Zまたは単独で表示 することちでさる。 , A symbol, such as "Normal value", is displayed together with the urine glucose concentration in Z or alone.
[0017] また、 pH測定器に無線または有線の通信機能を付加することにより、ノソコンへの データ格納、インターネット、携帯電話などを用いた病院への連絡なども可能となる。  [0017] In addition, by adding a wireless or wired communication function to the pH measuring device, it becomes possible to store data in a computer, contact the hospital using the Internet, a mobile phone, or the like.
[0018] 実施例 1 Example 1
グルコースォキシダーゼ (シグマ社製品 G-7016) 1325単位、フェリシアン化カリウム（ 和光純薬工業製品) 28mgを純水 lmlに溶カゝした水溶液を調製し、この水溶液 100 1を 、 I X lcmのろ紙 (東洋濾紙製品定性ろ紙 No.1)に滴下して浸み込ませ、冷蔵庫で一 晚乾燥させて、担体とした。  An aqueous solution was prepared by dissolving 1325 units of glucose oxidase (product of Sigma G-7016) and 28 mg of potassium ferricyanide (product of Wako Pure Chemical Industries) in 1 ml of pure water. Filter paper product qualitative filter paper No.1) was dropped and infiltrated, and dried in a refrigerator for a while to obtain a carrier.
[0019] コントロール尿 (バイオ'ラッドラボラトリーズ社製品 Quantitative Urine Control "Nomal")に 19.2、 28.6、 48.1、 53、 98および 200.8mg/dlとなるようにグルコース (シグ マ社製品)を加え、異なるグルコース濃度の尿標準サンプルを作成した。なお、すべ ての尿標準サンプルの初期 PHは、 6.1であった。 [0019] Control urine (Bio'Rad Laboratories product Quantitative Urine Control Glucose (Sigma product) was added to "Nomal") at 19.2, 28.6, 48.1, 53, 98 and 200.8 mg / dl to prepare urine standard samples with different glucose concentrations. It should be noted that the initial P H of urine standard sample of all and was 6.1.
[0020] pH測定機器として、底部に pH電極が設置されて ヽる簡易 pHメーター (堀場製作所 製品 Twin pH B-211)を用い、このサンプル採取部分 (底部)に、上記担体を設置し、 尿標準サンプルを 400 1滴下した後、 pH変化を 30(參)、 60(H), 120(D), 180(Δ), 240( Χ)、 300(〇)秒ごとに記録した。得られた結果は、図 1に示される。このグラフより 、 120秒の測定値で検量性がみられることが確認された。  [0020] As a pH measuring instrument, a simple pH meter (Twin pH B-211 manufactured by HORIBA, Ltd.) having a pH electrode at the bottom was used. After dropping 400 1 of the standard sample, the pH change was recorded every 30 (see), 60 (H), 120 (D), 180 (Δ), 240 (Χ), and 300 (〇) seconds. The results obtained are shown in FIG. From this graph, it was confirmed that calibration was observed at the measured value of 120 seconds.
[0021] 実施例 2  Example 2
実施例 1にお 、て、グルコースォキシダーゼおよびフェリシアンィ匕カリウムの試薬量 力 倍に変更されて用いられた。得られた結果は、図 2に示され、 60秒後の測定値 で検量性がみられることが確認された。  In Example 1, the reagents of glucose oxidase and potassium ferricyanide were used in a volume change of 1: 1. The obtained results are shown in Fig. 2, and it was confirmed that the measured values after 60 seconds showed calibration.
[0022] 実施例 3 Example 3
実施例 1にお 、て、グルコースォキシダーゼおよびフェリシアンィ匕カリウムの試薬量 力 ^倍に変更されて用いられた。得られた結果は、図 3に示され、 30秒後の測定値で 検量性がみられることが確認された。  In Example 1, the reagents of glucose oxidase and potassium ferricyanide were used in a state where the reagent volumes were changed to 倍 times. The obtained results are shown in Fig. 3, and it was confirmed that the measured values after 30 seconds showed calibration.
[0023] 実施例 4 Example 4
実施例 1にお 、て、グルコースォキシダーゼおよびフェリシアンィ匕カリウムの試薬量 力^倍に変更されて用いられた。得られた結果は、図 4に示され、全体として 60秒後に は濃度毎のはっきりとした差が現われ、尿糖濃度を測定できることが判った。検量範 囲は、約 50から 200mgZdlであった。  In Example 1, the reagents used for glucose oxidase and potassium ferricyanide were changed to twice the reagent capacity. The results obtained are shown in FIG. 4, and a clear difference between the concentrations appears after 60 seconds as a whole, indicating that the urinary glucose concentration can be measured. The calibration range was approximately 50 to 200 mg Zdl.
[0024] 実施例 5 Example 5
実施例 1において、担体の代わりにグルコースォキシダーゼ (G-7016)3975単位、フ エリシアンィ匕カリウム 84mgを紅茶用パック (リプトン社製品)に封入したものが用いられ た。得られた結果は、図 5に示され、全体として 60秒後には濃度ごとのはっきりとした 差が現われ、尿糖濃度を測定できることが判った。検量範囲は約 30から 200mgZdlで めつに。  In Example 1, instead of the carrier, a product in which 3975 units of glucose oxidase (G-7016) and 84 mg of potassium potassium were used in a black tea pack (Lipton) was used. The obtained results are shown in FIG. 5, and a clear difference appeared for each concentration after 60 seconds as a whole, indicating that the urinary glucose concentration can be measured. Calibration range is about 30 to 200mgZdl.

Claims

請求の範囲 The scope of the claims
[1] pH測定器の pH電極部またはその付近に、グルコースォキシダーゼまたはダルコ一 スデヒドロゲナーゼとメディエーターを含浸させた担体、あるいはグルコースォキシダ ーゼまたはグルコースデヒドロゲナーゼとメディエーターを封入した袋体力 なる反応 体を設置した尿糖バイオセンサ。  [1] A carrier in which glucose oxidase or dalcos dehydrogenase and a mediator are impregnated in or near a pH electrode part of a pH meter, or a bag-like reactant in which glucose oxidase or glucose dehydrogenase and a mediator are sealed. A urine glucose biosensor equipped with.
[2] グルコースォキシダーゼまたはグルコースデヒドロゲナーゼとメディエーターと共に 乾燥剤が用いられた請求項 1記載の尿糖バイオセンサ。 [2] The urine glucose biosensor according to claim 1, wherein a desiccant is used together with glucose oxidase or glucose dehydrogenase and a mediator.
[3] 担体または袋体が紙または不織布である請求項 1記載の尿糖バイオセンサ。 [3] The urine glucose biosensor according to claim 1, wherein the carrier or the bag is paper or nonwoven fabric.
[4] メディエーターがフェリシアン化カリウム、フエ口センまたはパラべンゾキノンである請 求項 1記載の尿糖バイオセンサ。 [4] The urinary glucose biosensor according to claim 1, wherein the mediator is potassium ferricyanide, fuessen or parabenzoquinone.
[5] pH測定器の表示が尿糖濃度および Zまたは尿糖濃度に対応した文章、単語、図 形、記号である請求項 1記載の尿糖バイオセンサ。 5. The urine glucose biosensor according to claim 1, wherein the indication on the pH meter is a sentence, word, figure, or symbol corresponding to the urine sugar concentration and Z or urine sugar concentration.
[6] pH測定器として、尿の初期 pHに基づく検量線を記憶させたものが用いられた請求 項 1記載の尿糖バイオセンサ。 [6] The urine glucose biosensor according to claim 1, wherein a pH meter that stores a calibration curve based on the initial pH of urine is used.
[7] 水溶液としたダルコースォキシダーゼまたはダルコ一スデヒドロゲナーゼとメデイエ 一ターを担体に滴下した後、乾燥させて含浸することを特徴とする請求項 1記載の尿 糖バイオセンサの製造方法。 7. The method for producing a urinary glucose biosensor according to claim 1, wherein the aqueous solution of dalcosoxidase or dalcos dehydrogenase and the mediator are dropped on a carrier, and then dried and impregnated.
[8] pH電極がガラス電極またはイオン感応性電界効果型トランジスターである請求項 1 記載の尿糖バイオセンサ。 [8] The urine glucose biosensor according to claim 1, wherein the pH electrode is a glass electrode or an ion-sensitive field-effect transistor.
[9] pH測定器に無線または有線の通信機能が付加された請求項 1記載の尿糖バイオ センサ。 [9] The urine glucose biosensor according to claim 1, wherein a wireless or wired communication function is added to the pH meter.
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7741069B2 (en) 2005-12-27 2010-06-22 Canon Kabushiki Kaisha Mesoporous material having dendritic skeleton containing immobilized glucose dehydrogenase

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JPH10142193A (en) * 1996-11-13 1998-05-29 Toa Denpa Kogyo Kk Method and apparatus for measuring concentration with utilizing phchange reaction
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US3707455A (en) * 1968-07-15 1972-12-26 Ibm Measuring system
JPS55118850A (en) * 1979-03-06 1980-09-12 Fujishima Daishiro Sheet to which chemical is fixed
JPS59104933A (en) * 1982-12-08 1984-06-18 三菱瓦斯化学株式会社 Filling sheet and its manufacture
JPH10142193A (en) * 1996-11-13 1998-05-29 Toa Denpa Kogyo Kk Method and apparatus for measuring concentration with utilizing phchange reaction
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