JPS5861459A - Analyzing device for creatine and creatinine - Google Patents
Analyzing device for creatine and creatinineInfo
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- JPS5861459A JPS5861459A JP56158780A JP15878081A JPS5861459A JP S5861459 A JPS5861459 A JP S5861459A JP 56158780 A JP56158780 A JP 56158780A JP 15878081 A JP15878081 A JP 15878081A JP S5861459 A JPS5861459 A JP S5861459A
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- 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
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Abstract
Description
【発明の詳細な説明】
本発明は血液や尿などの体液中のクレアチンとクレアチ
ニンの分析装置に関する。DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an apparatus for analyzing creatine and creatinine in body fluids such as blood and urine.
血液および尿中のクレアチンとクレアチニンの濃度は腎
臓機能の診断指標として重要な臨床検査項目とされてい
る。従来、フレア）とを分析するにはピクリン酸のアル
カリ性溶液中での発色反応を利用するＪ’ａｆｆｅ’反
応が用いられ、クレアチンは試料に酸を加えて加熱脱水
することによってりレアチニンに変化させてから同様に
分析していた。The concentrations of creatine and creatinine in blood and urine are considered important clinical test items as diagnostic indicators of kidney function. Conventionally, the J'affe' reaction, which utilizes a color reaction in an alkaline solution of picric acid, has been used to analyze creatine (flare), and creatine is converted to reatinine by adding acid to the sample and heating and dehydrating it. I then analyzed it in the same way.
しかし、Ｊａｆｆｅ’反応は必ずしもクレアチニンに特
異的ではなく他の物質とも反応する可能性があるので、
クレアチニン濃度を正確に分析することは困難とされて
いた。However, the Jaffe' reaction is not necessarily specific to creatinine and may react with other substances.
It has been difficult to accurately analyze creatinine concentration.
近年になって酵素試薬を用いるクレアチニンの分析法が
いくつか報告されている。例えば森下氏等はクレアチニ
ンディミナーゼ（Ｅ、 Ｃ，３，５，４゜２）溶液とア
ンモニアガス電極を用いる方法を提安している（衛生検
査、２８，１７４．１９６９）。In recent years, several analytical methods for creatinine using enzyme reagents have been reported. For example, Mr. Morishita et al. proposed a method using a creatinine diminase (E, C, 3, 5, 4°2) solution and an ammonia gas electrode (Sanitary Inspection, 28, 174, 1969).
また、Ｍａｙｅｒｈｏｆｆ氏等はアンモニア電極の先端
に膜を包んだ酵素溶液を装着する酵素電極を使用する方
法を提案している（ Ａｎａｌ、 Ｃｈｉｍ、 Ａｃｒ
ａ８５．２７７．１９７６）カＳ１ これらはいず九も
日常分析法としては採用されていない。In addition, Mayerhoff et al. have proposed a method using an enzyme electrode in which an enzyme solution wrapped in a membrane is attached to the tip of an ammonia electrode (Anal, Chim, Acr.
a85.277.1976) KaS1 None of these methods have been adopted as routine analytical methods.
本発明は生体液中のクレアチンとクレアチニンを同時に
分析できる簡便な分析装置を提供することを目的とし、
その特徴とするところは、クレアチンアミジノヒドロラ
ーゼとザルコシンオキシダーゼを固定化した酵素膜を感
応面に装着した第１の酵素電極と、クレアチニンアミド
ヒドロラーゼとクレアチンアミジノヒドロラーゼとザル
コシンオキシダーゼを固定化した酵素膜を感応面に装着
した第２の酵素電極とを設置し、第１の酵素電極の出力
よりクレアチンの濃度を求め、第２の酵素電極と第１の
酵素電極の出力差よりクレアチニンの濃度を求めるごと
く構成したことにある。The purpose of the present invention is to provide a simple analyzer that can simultaneously analyze creatine and creatinine in biological fluids.
Its features include the first enzyme electrode, which has an enzyme membrane on which creatinine amidinohydrolase and sarcosine oxidase are immobilized, and the enzyme membrane with which creatinine amidinohydrolase, creatine amidinohydrolase, and sarcosine oxidase are immobilized. A second enzyme electrode attached to the sensitive surface is installed, the concentration of creatine is determined from the output of the first enzyme electrode, and the concentration of creatinine is determined from the difference in output between the second enzyme electrode and the first enzyme electrode. The reason lies in the fact that it is structured as follows.
第１図は本発明の一実施例である分析装置の系統図であ
る。試料溶液１１を収容した測定容器１９には第１酵素
電極１と第２酵素電極３とが垂直に挿入されており、第
１酵素電極１の下端は次の物質を含む感応膜で封止され
ている。FIG. 1 is a system diagram of an analyzer that is an embodiment of the present invention. A first enzyme electrode 1 and a second enzyme electrode 3 are vertically inserted into a measurement container 19 containing a sample solution 11, and the lower end of the first enzyme electrode 1 is sealed with a sensitive membrane containing the following substances. ing.
クレアチンアミジノヒドロラーゼ（Ｅ、 Ｃ，３，５゜
３．３．以後ＣＩと記す）とザルコシンオキシダーゼ（
Ｅ、 Ｃ，１，５，３，１，以後ＳＯＤ、Ｊｅ−ｔ’）
を含む感応膜で、以後Ｃｌ−８ＯＤ膜２と記す。Creatine amidinohydrolase (E, C, 3,5°3.3.hereinafter referred to as CI) and sarcosine oxidase (
E, C, 1, 5, 3, 1, hereafter SOD, Je-t')
This is a sensitive film containing Cl-8OD film 2, hereinafter referred to as Cl-8OD film 2.
また、第２酵素電極３の下端は次の物質を含む感応膜で
封止されている。Further, the lower end of the second enzyme electrode 3 is sealed with a sensitive membrane containing the following substance.
上記ＣＩとＳＯＤにクレアチニンアミドヒドロラーゼ（
Ｅ、 Ｃ，３，５，２，ｎ、以後ＣＮと記す）を追加し
た感応膜で、以後ＣＮ−Ｃｌ−８ＯＤ膜４と記す。Creatinine amide hydrolase (
E, C, 3, 5, 2, n, hereinafter referred to as CN), and hereinafter referred to as CN-Cl-8OD film 4.
なお、これらの感応膜は次のような酵素接触反応を行う
。Note that these sensitive membranes perform the following enzyme contact reaction.
クレアチニン＋Ｈ２ＯＣＮクレアチン ・・曲（１）ク
レアチン＋Ｈ２０ＣＮ尿素＋ザルコシン・・・（２）ザ
ルコシン＋０２＋Ｈ２０□ＳＯＤホルムアルデヒド＋グ
リシン＋Ｈ２０・・・・・・・・・・・・・・・（３）
即ち、（２）、 （３）の反応を利用して（３）式の０
□を検出してクレアチンを求める。なお、この場合は酸
素電極を用いて０２を検出しているが、過酸化水素電極
を用いた時はＨ２０□を検出することになる。Creatinine + H2OCN Creatine... Song (1) Creatine + H20CN Urea + Sarcosine... (2) Sarcosine + 02 + H20 SOD Formaldehyde + Glycine + H20... (3)
That is, by using the reactions (2) and (3), 0 in equation (3)
Detect □ to determine creatine. In this case, 02 is detected using an oxygen electrode, but when a hydrogen peroxide electrode is used, H20□ is detected.
また、クレアチニンの分析は（１）〜（３）式によりＣ
Ｎ、ＣＩ、ＳＯＤ酵素を利用し、クレアチンと同じ検出
法で分析することができる。なお、検出器としては上記
のごとく酸素電極および過酸化水素電極を用いることが
感応膜との組合わせ上適当で、第１酵素電極１１第２酵
素電極３はいずれかの電極を内蔵している。In addition, the analysis of creatinine is performed using formulas (1) to (3).
It can be analyzed using the same detection method as creatine using N, CI, and SOD enzymes. As a detector, it is appropriate to use an oxygen electrode and a hydrogen peroxide electrode as described above in combination with a sensitive membrane, and the first enzyme electrode 11 and the second enzyme electrode 3 have either one of them built-in. .
このように構成された第１酵素電極１の出力は第１増幅
器１５に送られて増幅され演算器１７に入力される。同
様に第２酵素電極３の出力は第２増幅器１６に送られて
増幅され演算器１７に入る。The output of the first enzyme electrode 1 configured as described above is sent to the first amplifier 15, amplified, and input to the arithmetic unit 17. Similarly, the output of the second enzyme electrode 3 is sent to the second amplifier 16, amplified, and input to the arithmetic unit 17.
この演算器１７では所定の演算を行ってクレアチンおよ
びクレアチニンの分析値を同時に表示する。This calculator 17 performs predetermined calculations and simultaneously displays the analytical values of creatine and creatinine.
上記は測定系であるが、この分析操作を円滑に行うため
には次のような操作系が用いられている。Although the above is a measurement system, the following operation system is used to smoothly perform this analysis operation.
即ち、感応膜であるＣｌ−８ＯＤ膜２．ＣＮ−Ｃｌ−８
ＯＤ膜４に新鮮な試料溶液１１を常に接触させるだめの
スターク−１０を用い、その攪拌子９を測定容器１９中
に設置している。また、血清・尿等の試料５と緩衝溶液
６の吸引管を分注器７に接続し、分注器７の吐出ノズル
８は測定容器１９内に開口させている。一方、測定終了
した排液１４は測定容器１９の試料溶液１１内に先端を
挿入した吸引ノズル１２よりジッパ１３を作動されるこ
とによって排液容器内に、排出される。That is, the Cl-8OD film 2. which is a sensitive film. CN-Cl-8
A Stark-10 is used to keep the fresh sample solution 11 in contact with the OD membrane 4, and its stirrer 9 is placed in the measurement container 19. Further, suction tubes for a sample 5 such as serum or urine and a buffer solution 6 are connected to a dispenser 7, and a discharge nozzle 8 of the dispenser 7 is opened into a measurement container 19. On the other hand, the drained liquid 14 after the measurement is discharged into the drained liquid container by operating the zipper 13 from the suction nozzle 12 whose tip is inserted into the sample solution 11 of the measurement container 19.
この分析装置による分析法の概略を次に説明する。試料
５とその緩衝溶液６は分注器７によって測定容器１９中
に送られ、スターラー１０を作動させて攪拌子９を回転
させることによって常に新鮮な液をＣ１，−８ＯＤ膜２
やＣＮ−Ｃｌ−８ＯＤ膜４に接触させる。この時試料中
のクレアチニンは酵素ＣＩおよびＳＯＤの接触作用で分
解され、（３）式によって感応膜中の酸素濃度は減少し
、過酸化水素が生成する。したがって、指示電極として
酸素電極を用いる場合は、第１酸素電極はクレアチン濃
度に関係する負の信号が得られ、また、過酸化水素電極
を用いればクレアチン濃度に関する正の信号が得られる
ので、これらの内いずれかの出力信号を増幅器１６に導
いて増幅した後、演算器１７で濃度換算し、表示器１８
にクレアチン濃度として表示する。An outline of the analysis method using this analyzer will be explained next. The sample 5 and its buffer solution 6 are sent into the measurement container 19 by the dispenser 7, and by operating the stirrer 10 and rotating the stirrer 9, a fresh solution is constantly supplied to the C1, -8OD membrane 2.
or CN-Cl-8OD film 4. At this time, creatinine in the sample is decomposed by the contact action of enzymes CI and SOD, the oxygen concentration in the sensitive membrane decreases according to equation (3), and hydrogen peroxide is produced. Therefore, when an oxygen electrode is used as the indicator electrode, the first oxygen electrode provides a negative signal related to the creatine concentration, and when a hydrogen peroxide electrode is used, a positive signal related to the creatine concentration is obtained. After guiding the output signal of one of them to the amplifier 16 and amplifying it, the arithmetic unit 17 converts it into concentration, and displays it on the display 18.
Displayed as creatine concentration.
一方、ＣＮ−Ｃｌ−８ＯＤ膜４を装着した第２酵素電極
３では（１）〜（３）式によりクレアチニンが分解され
るが、同時にクレアチンも（２）、 （３）式によって
分解される。したがって、第２酵素電極３ではクレアチ
ンとクレアチニ／の合量に関する信号が得られるので、
この出力信号を増幅した後演算器１７にてはこの信号と
第１酵素電極ｌの信号との差をクレアチニン濃度と定め
て表示する。On the other hand, in the second enzyme electrode 3 equipped with the CN-Cl-8OD membrane 4, creatinine is decomposed according to equations (1) to (3), and at the same time, creatine is also decomposed according to equations (2) and (3). Therefore, the second enzyme electrode 3 can obtain a signal regarding the total amount of creatine and creatinine.
After amplifying this output signal, the arithmetic unit 17 determines the difference between this signal and the signal from the first enzyme electrode 1 as the creatinine concentration and displays it.
第２図は第１図の分析装置による検量線図で、横軸はク
レアチン又はクレアチニンの濃度をｍｇ／ ｄ ／＝で
示し、縦軸にはそれらの成分のピーク高さを示している
。この場合の試料を希釈する緩衝溶液にはリン酸緩衝溶
液ｐＨ７，５を用い、固定化酵素膜は第６図（Ｂ）に示
している２層製造のものを過酸化水素電極に装着して使
用した。FIG. 2 is a calibration curve diagram obtained by the analyzer shown in FIG. 1, in which the horizontal axis shows the concentration of creatine or creatinine in mg/d/=, and the vertical axis shows the peak heights of these components. In this case, a phosphate buffer solution pH 7.5 was used as the buffer solution for diluting the sample, and the immobilized enzyme membrane was a two-layered one shown in Figure 6 (B) and attached to the hydrogen peroxide electrode. used.
なお、この検量線を作成するに当って予めザルコシンの
溶液を緩衝溶液の流れに注入し、第１酵素雷極１と第２
酵素電極２の出力感度の比較を行い、クレアチニン濃度
を分析する場合は第１酵素電極の出力を校正した後で差
を求めた。第２図に示すごとくクレアチンはクレアチニ
ンよりも高感度で検出されるが、共に良好な比例性を示
している。In addition, when creating this calibration curve, a solution of sarcosine was injected into the flow of the buffer solution in advance, and the first enzyme lightning electrode 1 and the second
The output sensitivity of enzyme electrode 2 was compared, and when analyzing creatinine concentration, the difference was determined after calibrating the output of the first enzyme electrode. As shown in FIG. 2, creatine is detected with higher sensitivity than creatinine, but both show good proportionality.
本実施例の分析装置は、測定容器に収容された試料溶液
中にＣｌ−８ＯＤ膜を取付けた第１酵素電極とＣＮ−Ｃ
ｌ−８ＯＤ膜を取付けた第２酵素電極を浸漬して試料溶
液を攪拌し、各酵素電極の出力信号を増幅・演算処理し
て表示することにより、試料中のクレアチン、クレアチ
ニンの濃度を同時に高精度に分析することができるとい
う効果が得られる。The analyzer of this example consists of a first enzyme electrode equipped with a Cl-8OD film and a CN-C
The concentration of creatine and creatinine in the sample can be simultaneously increased by immersing the second enzyme electrode with the l-8OD membrane attached, stirring the sample solution, and amplifying, calculating and displaying the output signals of each enzyme electrode. The effect of being able to analyze with precision is obtained.
第１図の装置はディスクリート方式の分析装置であり微
小量の試料でも測定可能であるが、試料５を緩衝溶液６
で希釈しているので低濃度試料の場合はＳ／Ｎ比が低下
することがアリ、十分な分析精度は得られない。これを
改善したのが次の実施例である。The device shown in Figure 1 is a discrete analyzer and is capable of measuring even minute amounts of samples.
Since the sample is diluted with water, the S/N ratio may decrease in the case of a low-concentration sample, making it impossible to obtain sufficient analysis accuracy. The following embodiment improves this problem.
第３図は本発明の他の実施例である分析装置の系統図で
あり、第１図と同じ部分には同一符号を付しである。こ
の場合はフロ一方式を採用しており、キャリアー溶液２
０を送液ポンプ２１で吸引圧送して第１酵素電極１を収
容する第１フローセル２３ａと、第２酵素電極３を収容
する第２フローセル２３ｂとで構成されている。なお、
キャリアー溶液２０は第１図の場合の緩衝溶液を使用す
ることができる。FIG. 3 is a system diagram of an analyzer according to another embodiment of the present invention, in which the same parts as in FIG. 1 are given the same reference numerals. In this case, a one-flow system is used, and the carrier solution
The first flow cell 23a accommodates the first enzyme electrode 1 and the second flow cell 23b accommodates the second enzyme electrode 3. In addition,
As the carrier solution 20, the buffer solution shown in FIG. 1 can be used.
試料５は試料注入部２２に注入されてキャリアー溶液２
０の流れに乗り、まず第１フローセル２３ａに運ばれる
。第１フローセル２３ａにはＣｌ−８ＯＤ膜２が装着さ
れた第１酵素市極１が設置されており、Ｃｌ−８０１）
膜２は試料流に接触する。したがって、第１酵素電極１
ではクレアチン濃度に関する信号が得られる。The sample 5 is injected into the sample injection part 22 and the carrier solution 2
0 flow and is first carried to the first flow cell 23a. In the first flow cell 23a, a first enzyme market 1 equipped with a Cl-8OD membrane 2 is installed, and a Cl-801)
Membrane 2 is in contact with the sample stream. Therefore, the first enzyme electrode 1
will give you a signal regarding creatine concentration.
この試料５を含むキャリアー溶液２００層は第２フロー
セル２３ｂに導入される。この第２フローセル２３ｂに
はＣＮ−Ｃｌ−８ＯＤ膜４を装着した第２酵素雷極３が
設置されており、クレアチンとクレアチニンの含量に関
係する信号が得られる。第１酵素電極１と第２酵素電極
３の出力信号は夫々増幅され、演算器１７で第１酵素電
極１の出力はクレアチン濃度に、また、第１酵素電極と
第２酵素電極の出力信号の差がクレアチニン濃度に換算
され、夫々の値が表示器１８に表示される。A carrier solution 200 layer containing this sample 5 is introduced into the second flow cell 23b. A second enzyme lightning electrode 3 equipped with a CN-Cl-8OD membrane 4 is installed in the second flow cell 23b, and a signal related to the contents of creatine and creatinine can be obtained. The output signals of the first enzyme electrode 1 and the second enzyme electrode 3 are each amplified, and the output of the first enzyme electrode 1 is converted into the creatine concentration by the arithmetic unit 17, and the output signals of the first enzyme electrode and the second enzyme electrode are The difference is converted into a creatinine concentration, and each value is displayed on the display 18.
第４図は第３図の分析装置の記録線図であり、第１酵素
電極１の出力をＡ、第２酵素電極３の出力をＢとすると
、この２つのピークは完全に分離しており、均斉のとれ
たピーク形状を示している。Figure 4 is a recording diagram of the analyzer shown in Figure 3. If the output of the first enzyme electrode 1 is A and the output of the second enzyme electrode 3 is B, these two peaks are completely separated. , showing a well-balanced peak shape.
また、一度の異なる数種の試料を分析した時は、第２図
と同様にそのピーク高さは良好な直線性が得られること
を確認した。Furthermore, when several different types of samples were analyzed at the same time, it was confirmed that the peak heights had good linearity as shown in FIG. 2.
本実施例の分析装置は、一対のフローセルを連通させ、
第１のフローセルにはＣｌ−８ＯＤ膜を取付けた第１酵
素電極を設置し、第２のフローセルにはＣＮ−Ｃｌ−８
ＯＤ膜を取付けた第２酵素電極を設置して試料成分を測
定することによって、極めて小量の試料でも迅速正確に
分析できるという効果が得られる。The analyzer of this example communicates a pair of flow cells,
A first enzyme electrode equipped with a Cl-8OD membrane was installed in the first flow cell, and a CN-Cl-8 OD membrane was installed in the second flow cell.
By installing a second enzyme electrode equipped with an OD membrane and measuring sample components, it is possible to quickly and accurately analyze even an extremely small amount of sample.
第５図は第３図の変形例であるフローセルの説明図で、
この場合は第１酵素電極１と第２酵素電極３を１個のフ
ローセル２４に並設しである。このようにすればフロー
セル２４の設置場所が節約できると共に流路２５が短縮
されるので、更に短時間で分析できるという利点が得ら
れる。FIG. 5 is an explanatory diagram of a flow cell that is a modification of FIG.
In this case, the first enzyme electrode 1 and the second enzyme electrode 3 are arranged side by side in one flow cell 24. In this way, the installation space of the flow cell 24 can be saved and the flow path 25 can be shortened, so that an advantage can be obtained that analysis can be performed in a shorter time.
上記のＣｌ−８ＯＤ膜２堂ＣＮ−Ｃｌ−８ＯＤ−膜４等
の酵素膜を作成する時は、例えば多孔性膜内に各酵素を
注入して膜母材と架橋結合させる方法、酵素溶液を２枚
の多孔性膜内に封し込める方法、多孔性膜表面にゲル状
の酵素を塗布する方法等各種の酵素膜製法が適用できる
。また、上記のごとく複数種の酵素を一枚の膜内に固定
化する場合は、次のような方法も用いられる。When creating enzyme membranes such as the above-mentioned Cl-8OD membrane 2 and CN-Cl-8OD-membrane 4, there are methods, for example, injecting each enzyme into a porous membrane and crosslinking it with the membrane base material, or using an enzyme solution. Various enzyme membrane manufacturing methods can be applied, such as a method in which the enzyme is sealed within two porous membranes, and a method in which a gel-like enzyme is applied to the surface of the porous membrane. Furthermore, when multiple types of enzymes are immobilized within a single membrane as described above, the following method may also be used.
第６図は複数種の酵素を一枚の酵素膜内に固定化した状
態の説明図で、第６図（Ａ）はＣＮ、ＣＩ。Figure 6 is an explanatory diagram of a state in which multiple types of enzymes are immobilized within a single enzyme membrane, and Figure 6 (A) shows CN and CI.
ＳＯＤ等を混合して膜母材に一様に含ませて固定化した
もの、第６図（Ｂ）は２層よりなる酵素膜で、例えば試
料に接する側にはＣＮとＣＩの酵素を混合して固定化し
、電極に接する側にはＳＯＤを固定した場合は好結果が
得られる。第４図（（Ｊは３層構造の酵素膜で、試料に
接する側にはＣＮ、中間層にはＣＩ、電極側にはＳＯＤ
を固定化したものである。これはＣＮ−Ｃｌ−８ＯＤ膜
４の場合であるが、Ｃｌ−８ＯＤ膜２０時は、第４図（
Ａ）。Figure 6 (B) is an enzyme membrane consisting of two layers, in which SOD, etc. is mixed and uniformly impregnated in the membrane base material, and for example, CN and CI enzymes are mixed on the side that contacts the sample. Good results can be obtained if SOD is immobilized on the side in contact with the electrode. Figure 4 (J is a three-layered enzyme membrane, with CN on the side in contact with the sample, CI on the middle layer, and SOD on the electrode side.
is fixed. This is the case of CN-Cl-8OD film 4, but in the case of Cl-8OD film 20, as shown in Fig. 4 (
A).
第４図（Ｂ）の方法が採用できるし、第４図（Ｂ）の場
合は試料側にはＣ１１電極側にＳＯＤを固定化する。The method shown in FIG. 4(B) can be adopted, and in the case of FIG. 4(B), SOD is immobilized on the C11 electrode side on the sample side.
本発明のクレアチンとクレアチニンの分析装置は、ＣＩ
とＳＯＤを固定化した酵素膜を感応面に装着した第１酵
素電極と、ＣＩとＳＯＤとＣＮを固定化した酵素膜を感
応面に装着した第２酵素電極とが試料液と接触する際に
生じる出力信号を処理することにより、迅速正確にその
目的を達することができるという効果が得られる。The creatine and creatinine analyzer of the present invention is a CI
When the first enzyme electrode has a sensitive surface equipped with an enzyme membrane on which CI, SOD, and CN are immobilized, and the second enzyme electrode has a sensitive surface equipped with an enzyme membrane on which CI, SOD, and CN are immobilized, when they come into contact with the sample solution. By processing the resulting output signals, the advantage is that the objective can be achieved quickly and precisely.
第１図は本発明の一実施例である分析装置の系統図、第
２図は第１図の分析装置による検量線図、第３図は本発
明の他の実施例である分析装置の系統図、第４図は第３
図の分析装置の記録線図、第５図は第３図の変形例であ
るフローセルの説明図、第６図は複数種の酵素を一枚の
酵素膜内に固定した状態の説明図である。
１・・・第１酵素電極、２・・・Ｃｌ−８ＯＤ膜、３・
・・第２酵素電極、４・・・ＣＮ−Ｃｌ−８ＯＤ膜、５
・・・試料、６・・・緩衝溶液、７・・・分注器、８・
・・吐出ノズ７ｔ、、９・・・攪拌子、１０・・・スタ
ーラー、１１・・・試料溶液、１２吸引ノズル、１３・
・・シラ、Ｃ１１４・・・排液、１５．１６・・・増幅
器、１７・・・演算器、１８・・・表示器、１９・・・
測定容器、２０・・・キャリアー溶液、２１・・・送液
ポンプ、２２・・・試料注入部、２３゜２４・・・フロ
ーセル、２５・・・流路。
代理人 弁理士 高橋明夫
冴 １ 図
業 ２０
笛ｌＡ目
一ン吟關
１！Ｊｊ口
鴬 ５ 図
′Ｒ６目Figure 1 is a system diagram of an analyzer that is an embodiment of the present invention, Figure 2 is a calibration curve diagram of the analyzer shown in Figure 1, and Figure 3 is a system diagram of an analyzer that is another embodiment of the present invention. Figure 4 is the third
Fig. 5 is an explanatory diagram of a flow cell which is a modification of Fig. 3; Fig. 6 is an explanatory diagram of a state in which multiple types of enzymes are immobilized within a single enzyme membrane. . 1... First enzyme electrode, 2... Cl-8OD membrane, 3...
...Second enzyme electrode, 4...CN-Cl-8OD membrane, 5
...Sample, 6.Buffer solution, 7.Dispenser, 8.
...Discharge nozzle 7t, 9... Stirrer, 10... Stirrer, 11... Sample solution, 12 Suction nozzle, 13...
...Silla, C114...Drainage, 15.16...Amplifier, 17...Arithmetic unit, 18...Display unit, 19...
Measurement container, 20...Carrier solution, 21...Liquid pump, 22...Sample injection part, 23°24...Flow cell, 25...Flow path. Agent Patent Attorney Akio Sae Takahashi 1 Business 20 Fue lAme Ichiin Ginkan 1! Jj Kuchiho 5 Figure'R6th
Claims (1)
装置において、クレアチンアミジノヒドロラーゼとザル
コシンオキシダーゼを固定化した酵素膜を感応面に装着
した第１の酵素電極と、クレアチニンアミドヒドロラー
ゼとクレアチンアミジノヒドロラーゼとザルコシンオキ
シダーゼを固定化した酵素膜を感応面に装着した第２の
酵素電極とを設置し、上記第１の酵素電極の出力より上
記クレアチンの濃度を求め、上記第２の酵素電極と上記
第１の酵素電極の出力差より上記クレアチニンの濃度を
求めるごとく構成したことを特徴とするクレアチンとク
レアチニンの分析装置。 ２、 上記第１の酵素電極と第２の酵素電極が、測定容
器中に並列して挿入され、その酵素膜を試料溶液に浸漬
している電極である特許請求の範囲第１項記載のクレア
チンとクレアチニンの分析装置。 ３、上記第１の酵素電極と第２の酵素電極が、直列に連
通させたフロルセルのキャリアーン溶液流路にその酵素
膜を浸漬して設置された電極である特許請求の範囲第１
項記載のクレアチンとクレアチニンの分析装置。
、 ４、上記酵素電極が、酵素電極である特許請求の範囲第
１項記載のクレアチンとクレアチニンの分析装置。 ５、上記酵素電極が、過酸化水素電極である特許請求の
範囲第１項記載のクレアチンとクレアチニンの分析装置
。[Scope of Claims] 1. A creatine and creatinine analyzer using an enzyme electrode, a first enzyme electrode having an enzyme membrane on which creatine amidinohydrolase and sarcosine oxidase are immobilized is mounted on the sensitive surface, and creatinine amidinohydrolase. and a second enzyme electrode whose sensitive surface is equipped with an enzyme membrane on which creatine amidinohydrolase and sarcosine oxidase are immobilized, the concentration of creatine is determined from the output of the first enzyme electrode, and the concentration of creatine is determined from the output of the first enzyme electrode. A device for analyzing creatine and creatinine, characterized in that it is configured to determine the concentration of creatinine from the output difference between the enzyme electrode and the first enzyme electrode. 2. The creatine according to claim 1, wherein the first enzyme electrode and the second enzyme electrode are electrodes that are inserted in parallel into a measurement container and whose enzyme membranes are immersed in a sample solution. and creatinine analyzer. 3. Claim 1, wherein the first enzyme electrode and the second enzyme electrode are electrodes whose enzyme membranes are immersed in a carrier solution flow path of a Florcell connected in series.
Creatine and creatinine analyzer as described in section.
4. The creatine and creatinine analyzer according to claim 1, wherein the enzyme electrode is an enzyme electrode. 5. The creatine and creatinine analyzer according to claim 1, wherein the enzyme electrode is a hydrogen peroxide electrode.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP56158780A JPS5861459A (en) | 1981-10-07 | 1981-10-07 | Analyzing device for creatine and creatinine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP56158780A JPS5861459A (en) | 1981-10-07 | 1981-10-07 | Analyzing device for creatine and creatinine |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS5861459A true JPS5861459A (en) | 1983-04-12 |
| JPH029302B2 JPH029302B2 (en) | 1990-03-01 |
Family
ID=15679164
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP56158780A Granted JPS5861459A (en) | 1981-10-07 | 1981-10-07 | Analyzing device for creatine and creatinine |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS5861459A (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6085359A (en) * | 1983-10-14 | 1985-05-14 | Matsushita Electric Works Ltd | Substance quantification |
| JPS6353461A (en) * | 1986-08-22 | 1988-03-07 | Japanese Res & Dev Assoc Bio Reactor Syst Food Ind | Measurement of peptide |
| EP0771867A3 (en) * | 1995-10-30 | 1998-09-02 | Ciba-Geigy Japan Limited | Enzyme electrode |
| AT409040B (en) * | 2000-08-11 | 2002-05-27 | Roche Diagnostics Gmbh | CREATININSENSOR CALIBRATION |
| JP2003533679A (en) * | 2000-05-16 | 2003-11-11 | エフ．ホフマン−ラ ロシュ アーゲー | Creatinine biosensor |
| JP2007524094A (en) * | 2004-02-16 | 2007-08-23 | ピー エー コンサルティング サービシーズ リミテッド | Analyte testing apparatus and method |
| EP2261647A1 (en) * | 2001-05-31 | 2010-12-15 | Instrumentation Laboratory Company | Cross-linked enzyme matrix and uses thereof |
| JP2021527194A (en) * | 2019-04-05 | 2021-10-11 | インストゥルメンテーション ラボラトリー カンパニー | How to improve the calibration accuracy of the creatinine / creatine sensor |
-
1981
- 1981-10-07 JP JP56158780A patent/JPS5861459A/en active Granted
Cited By (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0570102B2 (en) * | 1983-10-14 | 1993-10-04 | Matsushita Electric Works Ltd | |
| JPS6085359A (en) * | 1983-10-14 | 1985-05-14 | Matsushita Electric Works Ltd | Substance quantification |
| JPS6353461A (en) * | 1986-08-22 | 1988-03-07 | Japanese Res & Dev Assoc Bio Reactor Syst Food Ind | Measurement of peptide |
| EP0771867A3 (en) * | 1995-10-30 | 1998-09-02 | Ciba-Geigy Japan Limited | Enzyme electrode |
| JP2003533679A (en) * | 2000-05-16 | 2003-11-11 | エフ．ホフマン−ラ ロシュ アーゲー | Creatinine biosensor |
| JP4755802B2 (en) * | 2000-08-11 | 2011-08-24 | エフ．ホフマン−ラ ロシュ アーゲー | Calibration of creatinine sensor |
| WO2002014533A3 (en) * | 2000-08-11 | 2002-07-18 | Hoffmann La Roche | Creatinine sensor calibration |
| JP2004506224A (en) * | 2000-08-11 | 2004-02-26 | エフ．ホフマン−ラ ロシュ アーゲー | Calibration of creatinine sensor |
| US7371314B2 (en) | 2000-08-11 | 2008-05-13 | Roche Diagnostics Operations, Inc. | Creatinine sensor calibration |
| US7815788B2 (en) | 2000-08-11 | 2010-10-19 | Roche Diagnostics Operations, Inc. | Creatinine sensor calibration |
| AT409040B (en) * | 2000-08-11 | 2002-05-27 | Roche Diagnostics Gmbh | CREATININSENSOR CALIBRATION |
| EP2261647A1 (en) * | 2001-05-31 | 2010-12-15 | Instrumentation Laboratory Company | Cross-linked enzyme matrix and uses thereof |
| US8426192B2 (en) | 2001-05-31 | 2013-04-23 | Instrumentation Laboratory Company | Composite membrane containing a cross-linked enzyme matrix for a biosensor |
| US9388503B2 (en) | 2001-05-31 | 2016-07-12 | Instrumentation Laboratory Company | Cross-linked enzyme matrix and uses thereof |
| JP2007524094A (en) * | 2004-02-16 | 2007-08-23 | ピー エー コンサルティング サービシーズ リミテッド | Analyte testing apparatus and method |
| JP2021527194A (en) * | 2019-04-05 | 2021-10-11 | インストゥルメンテーション ラボラトリー カンパニー | How to improve the calibration accuracy of the creatinine / creatine sensor |
| JP2021528640A (en) * | 2019-04-05 | 2021-10-21 | インストゥルメンテーション ラボラトリー カンパニー | Compositions and methods for improved creatinine measurement accuracy and their use |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH029302B2 (en) | 1990-03-01 |
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