WO2005080953A1 - Biocapteur de glucose dans l’urine - Google Patents

Biocapteur de glucose dans l’urine 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
Authority
WO
WIPO (PCT)
Prior art keywords
glucose
urine
mediator
meter
biosensor according
Prior art date
Application number
PCT/JP2005/002391
Other languages
English (en)
Japanese (ja)
Inventor
Masao Gotoh
Hideaki Nakamura
Satoshi Koide
Fumiyo Kurusu
Isao Karube
Original Assignee
National Institute Of Advanced Industrial Science And Technology
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by National Institute Of Advanced Industrial Science And Technology filed Critical National Institute Of Advanced Industrial Science And Technology
Priority to JP2006510213A priority Critical patent/JP4247844B2/ja
Publication of WO2005080953A1 publication Critical patent/WO2005080953A1/fr

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Classifications

    • 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.

Abstract

Un biocapteur de glucose dans l’urine comprenant un pH-mètre et, placé sur une partie de l’électrode de pH ou dans les alentours de celle-ci, un réactif consistant en une poche contenant de la glucose oxydase ou de la glucose déshydrogénase et un médiateur, ou un support imprégné de glucose oxydase ou de glucose déshydrogénase et d’un médiateur. Lors de l’utilisation du biocapteur de glucose dans l’urine, les réactifs du support ou de la poche sont évacués chaque fois. Ainsi, même lorsque les réactifs sont stockés à l’état sec, toute modification structurelle des enzymes est légère, de ce fait stabilisant la performance du capteur. En conséquence, aussi bien le réactif que le pH-mètre est portatif, de sorte qu’à la différence des instruments de mesure classiques, il n’est pas nécessaire d’ajouter de tampon pour le stockage et la mesure de glucose dans l’urine peut être réalisée à tout moment et en tout lieu.
PCT/JP2005/002391 2004-02-25 2005-02-17 Biocapteur de glucose dans l’urine WO2005080953A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2006510213A JP4247844B2 (ja) 2004-02-25 2005-02-17 尿糖バイオセンサ

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2004049466 2004-02-25
JP2004-049466 2004-02-25

Publications (1)

Publication Number Publication Date
WO2005080953A1 true WO2005080953A1 (fr) 2005-09-01

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JP (1) JP4247844B2 (fr)
WO (1) WO2005080953A1 (fr)

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

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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 (ja) * 1982-12-08 1984-06-18 三菱瓦斯化学株式会社 充填シ−トおよびその製造法
JPH10142193A (ja) * 1996-11-13 1998-05-29 Toa Denpa Kogyo Kk ｐＨ変動反応を利用する濃度測定方法及びその装置
WO2002093151A1 (fr) * 2001-05-15 2002-11-21 Matsushita Electric Industrial Co., Ltd. Biocapteur

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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 (ja) * 1982-12-08 1984-06-18 三菱瓦斯化学株式会社 充填シ−トおよびその製造法
JPH10142193A (ja) * 1996-11-13 1998-05-29 Toa Denpa Kogyo Kk ｐＨ変動反応を利用する濃度測定方法及びその装置
WO2002093151A1 (fr) * 2001-05-15 2002-11-21 Matsushita Electric Industrial Co., Ltd. Biocapteur

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

Also Published As

Publication number Publication date
JPWO2005080953A1 (ja) 2007-10-25
JP4247844B2 (ja) 2009-04-02

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