JPS6288953A - Enzyme electrode - Google Patents
Enzyme electrodeInfo
- Publication number
- JPS6288953A JPS6288953A JP60230426A JP23042685A JPS6288953A JP S6288953 A JPS6288953 A JP S6288953A JP 60230426 A JP60230426 A JP 60230426A JP 23042685 A JP23042685 A JP 23042685A JP S6288953 A JPS6288953 A JP S6288953A
- Authority
- JP
- Japan
- Prior art keywords
- electrode
- enzyme
- membrane
- measurement
- electrolytic
- Prior art date
- Legal status (The legal status 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 status listed.)
- Pending
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- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Description
【発明の詳細な説明】
〔技術分野〕
この発明は、過酸化水素を検知物質とする酵素電極に関
する。DETAILED DESCRIPTION OF THE INVENTION [Technical Field] The present invention relates to an enzyme electrode using hydrogen peroxide as a detection substance.
過酸化水素を検知物質とする酵素電極は、一般に、測定
目的物質以外のアスコルビン酸等の還元性物質にも応答
性を示し、測定上、これらの還元性物質は、正の誤差を
与える。そのため、この正の誤差を除く方法として、酵
素電極とは別に、酵素のついていない補償極を用いて酵
素電極の測定値を補償極の測定値で補正する方法、ある
いは、アセチルセルロース膜の非対称膜を用い、膜の孔
径を利用して大きなサイズの分子（測定妨害物質）は透
過させないが１．過酸化水素を透過させるようにする方
法が用いられている。Enzyme electrodes that use hydrogen peroxide as a detection substance generally exhibit responsiveness to reducing substances such as ascorbic acid other than the measurement target substance, and these reducing substances give a positive error in measurement. Therefore, as a method to eliminate this positive error, there is a method to correct the measured value of the enzyme electrode using the measured value of the compensating electrode by using a compensating electrode that does not have an enzyme attached separately from the enzyme electrode, or a method using an asymmetric membrane of an acetyl cellulose membrane. The pore size of the membrane is used to prevent large molecules (measurement interfering substances) from passing through.1. A method is used that allows hydrogen peroxide to permeate.
しかしながら、前者の方法は、酵素電極とは別に新たに
補償極を設けなければならないという問題があり、後者
の方法は、非対称膜の作製が非常にむずかしく、非対称
膜の市販品は非常に高価であるという問題がある。However, the former method has the problem that a compensation electrode must be newly provided in addition to the enzyme electrode, and the latter method is extremely difficult to fabricate an asymmetric membrane, and commercially available asymmetric membranes are very expensive. There is a problem.
この発明は、このような事情に鑑みてなされたものであ
って、測定妨害物質除去膜を備えて、測定目的物質の測
定を正確に行え、そのうえ製造も簡単な、過酸化水素を
検知物質とする酵素電極を提供することを目的としてい
る。This invention was made in view of the above circumstances, and is a method that uses hydrogen peroxide as a detection substance, which is equipped with a measurement interfering substance removal membrane, allows accurate measurement of the measurement target substance, and is easy to manufacture. The purpose of this research is to provide an enzyme electrode that can
前記のような目的を達成するため、この発明は、過酸化
水素を検知物質とし、測定妨害物質除去膜を備えた酵素
電極であって、測定妨害物質除去膜として電解重合膜が
用いられていることを特徴とする酵素電極をその要旨と
している。以下に、この発明の詳細な説明する。In order to achieve the above object, the present invention provides an enzyme electrode that uses hydrogen peroxide as a detection substance and is equipped with a measurement interfering substance removal membrane, in which an electrolytic polymer membrane is used as the measurement interfering substance removal membrane. The gist of the invention is an enzyme electrode characterized by the following characteristics. The present invention will be explained in detail below.
この発明で用いられる、過酸化水素を検知物質とする酵
素電極としては、たとえば、グルコースオキシダーゼ等
の測定目的物質（基質）と反応して過酸化水素を生成す
る酵素が固定された酵素固定化膜が、白金板等の導電性
材料からなる電極本体表面に設けられたものがあげられ
る。The enzyme electrode that uses hydrogen peroxide as a detection substance used in this invention includes, for example, an enzyme-immobilized membrane on which an enzyme that reacts with a measurement target substance (substrate) such as glucose oxidase to generate hydrogen peroxide is immobilized. However, examples include those provided on the surface of an electrode body made of a conductive material such as a platinum plate.
グルコースオキシダーゼは、測定目的物質のグルコース
と反応して過酸化水素を生成させる。この酵素電極は、
たとえば、グルコースオキシダーゼ等の酵素、アルブミ
ンおよび架橋剤を含む酵素溶液を電極本体表面に塗布し
たのち、架橋反応を行って、膜を形成させるとともに膜
にゲルコールオキシダーゼ等の酵素を固定することによ
りつくることができる。ここで、アルブミンは、酵素固
定化膜のマトリックス成分となるものである。架橋剤と
しては、普通、グルタルアルデヒド等のジアルデヒドが
用いられる。Glucose oxidase reacts with glucose, which is the substance to be measured, to generate hydrogen peroxide. This enzyme electrode is
For example, an enzyme solution containing an enzyme such as glucose oxidase, albumin, and a crosslinking agent is applied to the surface of the electrode body, and then a crosslinking reaction is performed to form a membrane and an enzyme such as gelcol oxidase is immobilized on the membrane. be able to. Here, albumin serves as a matrix component of the enzyme-immobilized membrane. As the crosslinking agent, dialdehydes such as glutaraldehyde are commonly used.
この発明にかかる酵素電極は、前記のような酵素電極の
酵素固定化膜の外側に電解重合膜からなる測定妨害物質
除去膜を形成させることにより得ることができる。電解
重合膜は、たとえば、ピロール、チオフェン、アニリン
等の電解重合性物質を含む電解重合液に酵素電極を浸し
、電解処理を行うことにより形成することができる。電
解処理は、一般に、わずか数１０秒ですむので、電解重
合膜は、非常に簡単に形成させることができる。The enzyme electrode according to the present invention can be obtained by forming a measurement interfering substance removal membrane made of an electrolytic polymer membrane on the outside of the enzyme-immobilized membrane of the enzyme electrode as described above. The electrolytically polymerized membrane can be formed, for example, by immersing an enzyme electrode in an electrolytically polymerizing solution containing an electrolytically polymerizable substance such as pyrrole, thiophene, or aniline, and performing electrolytic treatment. Since the electrolytic treatment generally takes only a few tens of seconds, the electrolytic polymerized membrane can be formed very easily.
あらかじめ、電極本体に電解重合膜を設けておき、電解
重合膜の外側にグルコースオキシダーゼ等が固定された
酵素固定化膜を設けるようにしても、この発明にかかる
酵素電極を得ることができる。The enzyme electrode according to the present invention can also be obtained by providing an electrolytically polymerized membrane on the electrode body in advance, and then providing an enzyme-immobilized membrane on which glucose oxidase or the like is immobilized on the outside of the electrolytically polymerized membrane.
電解重合膜は、過酸化水素をよく透過させるが、測定妨
害物質をほとんど透過させない。したがって、この発明
にかかる酵素電極は、測定妨害物質の影響をほとんど受
けず、過酸化水素選択性を有し、補償極を設けなくても
測定目的物質の測定を非常に正確に行うことができる。Electrolytic polymer membranes allow hydrogen peroxide to pass through them well, but hardly any substances that interfere with measurement pass through them. Therefore, the enzyme electrode according to the present invention is almost unaffected by substances that interfere with measurement, has hydrogen peroxide selectivity, and can measure the target substance very accurately even without providing a compensation electrode. .
また、酵素固定化部よりも外側に電解重合膜を設けるよ
うにした場合等においては、電解重合膜は測定目的物質
の透過性がよいので、電解重合膜の代わりに、従来より
測定妨害物質除去膜として用いられているアセチルセル
ロース系の過酸化水素選択膜を用いた場合に比べて、一
般に、検出感度が高く、応答性も１５〜２０秒と迅速で
ある。In addition, in cases where an electrolytic polymer membrane is provided outside the enzyme immobilization area, electrolytic polymer membranes have good permeability to the measurement target substance, so they can be used instead of electrolytic polymer membranes to remove substances that interfere with measurement. Compared to the case where an acetyl cellulose-based hydrogen peroxide selective membrane is used as a membrane, the detection sensitivity is generally higher and the response is faster at 15 to 20 seconds.
つぎに、実施例および比較例について説明する実施例１
．２の酵素電極をつぎのようにしてつく っ た。Next, Example 1 to explain Examples and Comparative Examples
．． The second enzyme electrode was made as follows.
グルコースオキシダーゼ、アルブミンおよびグルタルア
ルデヒドを含む酵素溶液を白金電極に塗布して架橋反応
を行い、白金電極表面にグルコースオキシダーゼの固定
化膜を設けてグルコースオキシダーゼ（ＣＯＤ）電極を
つくった。つぎに、ピロールを０．２Ｍ、ＮａＣ１を０
．２Ｍの割合でそれぞれ含む電解重合液中にこの電極を
浸し、第１表に示されている条件で電解重合処理を行っ
て、電解重合膜を備えた酵素電極を得た。ただし、重合
電圧はＡｇ／ＡｇＣ１電極に対するものである。第１表
に示されているように、実施例１． ２の製造において
は、電解重合処理の重合時の電圧および重合時間（電解
時間）を変えるようにした。An enzyme solution containing glucose oxidase, albumin, and glutaraldehyde was applied to a platinum electrode to perform a crosslinking reaction, and a glucose oxidase immobilized membrane was provided on the surface of the platinum electrode to prepare a glucose oxidase (COD) electrode. Next, add pyrrole to 0.2M and NaCl to 0.
．． The electrodes were immersed in electrolytic polymerization solutions containing 2M each, and electrolytic polymerization was performed under the conditions shown in Table 1 to obtain enzyme electrodes equipped with electrolytic polymeric membranes. However, the polymerization voltage is for an Ag/AgC1 electrode. As shown in Table 1, Example 1. In the production of No. 2, the voltage and polymerization time (electrolysis time) during electrolytic polymerization were varied.
比較例の酵素電極は、電解重合膜を設けないようにした
ほかは、実施例１．２と同様にしてつくった。The enzyme electrode of the comparative example was produced in the same manner as in Example 1.2, except that the electrolytically polymerized membrane was not provided.
実施例１，２および比較例の酵素電極をｐＨ７，５のリ
ン酸緩衝液で洗浄したのち、グルコース測定用のフロー
システムに組み込み、グルコース、アスコルビン酸およ
び尿酸に対する応答感度を調べた。アスコルビン酸およ
び尿酸は測定妨害物質である。ただし、キャリアー流速
は３．’Ｏｍｌ／分、印加電圧は０．７Ｖ（対Ａｇ／Ａ
ｇＣ１電極）とし、サンプルとして、１００ｍｇ／ ｄ
ｉのグルコース溶液、１０ｍｇ／ ｄｉのアスコルビン
酸溶液および１０ｍｇ７ｄｌの尿酸溶液をそれぞれ１０
μｌずつ用いることとした。結果を第１表に示す。After washing the enzyme electrodes of Examples 1 and 2 and Comparative Example with a phosphate buffer solution of pH 7.5, they were incorporated into a flow system for measuring glucose, and the response sensitivity to glucose, ascorbic acid, and uric acid was examined. Ascorbic acid and uric acid are measurement interfering substances. However, the carrier flow rate is 3. 'Oml/min, applied voltage is 0.7V (vs. Ag/A
gC1 electrode), and as a sample, 100mg/d
i of glucose solution, 10 mg/di of ascorbic acid solution and 10 mg/di of uric acid solution, respectively.
It was decided to use μl each. The results are shown in Table 1.
第１表より、実施例１． ２の酵素電極は、測定妨害物
質に対して応答しないので、測定妨害物質が存在しても
グルコースを正確に測定することができるということが
わかる。これに対し、比較例は、測定妨害物質に対して
強く応答するので、測定妨害物質が存在するとグルコー
スを正確に測定することができないことがわかる。From Table 1, Example 1. It can be seen that the enzyme electrode No. 2 does not respond to measurement-interfering substances, so that glucose can be accurately measured even in the presence of measurement-interfering substances. On the other hand, the comparative example responds strongly to the substance interfering with measurement, which indicates that glucose cannot be measured accurately in the presence of the substance interfering with measurement.
つぎに、白金板に電解重合膜を設けて行った実験を説明
する。Next, an experiment conducted in which an electrolytically polymerized membrane was provided on a platinum plate will be explained.
前記と同様の電解重合液を用い、白金板に電解重合膜を
設けて、電極１〜３をっ（った。電解処理条件は第２表
に示されているとおりである。Using the same electrolytic polymerization solution as above, an electrolytic polymeric film was provided on a platinum plate, and electrodes 1 to 3 were formed.The electrolytic treatment conditions are as shown in Table 2.
電極１〜３および白金板を洗浄したのち、前記と同様の
フローシステムに組み込み、過酸化水素、アスコルビン
酸および尿酸に対する応答感度を調べた。測定条件は前
記と同じとし、サンプルとしては、６ｍｇ／ｄｉの過酸
化水素溶液、 １０ｍｇ／ｄｉのアスコルビン酸溶液
および１０ｍｇ／ｄｌの尿酸溶液をそれぞれ１０μｍず
つ用いることとした。結果を第２表に示す。After cleaning electrodes 1 to 3 and the platinum plate, they were installed in the same flow system as above, and the response sensitivity to hydrogen peroxide, ascorbic acid, and uric acid was examined. The measurement conditions were the same as above, and the samples were 10 μm each of a 6 mg/di hydrogen peroxide solution, a 10 mg/di ascorbic acid solution, and a 10 mg/dl uric acid solution. The results are shown in Table 2.
第２表より、電極１〜３は過酸化水素に対して応答する
が、アスコルビン酸や尿酸に対してはほとんど応答しな
いことがわかる。したがって、電極１〜３の表面に酵素
固定化膜を装着すれば、基質を正確に測定できる酵素電
極を得ることができる。Table 2 shows that electrodes 1 to 3 respond to hydrogen peroxide, but hardly respond to ascorbic acid or uric acid. Therefore, by attaching an enzyme-immobilized membrane to the surface of electrodes 1 to 3, an enzyme electrode capable of accurately measuring the substrate can be obtained.
なお、この発明で用いられる電解重合膜は、酵素電極に
設けるのではなく、単独にして、過酸化水素は透過する
がアスコルビン酸等の大きなサイズの分子は透過しない
過酸化水素選択膜として利用できる。The electropolymerized membrane used in this invention can be used alone, instead of being attached to an enzyme electrode, as a hydrogen peroxide selective membrane that allows hydrogen peroxide to pass through but not large molecules such as ascorbic acid. .
この発明にかかる酵素電極は、過酸化水素を検知物質と
し、測定妨害物質除去膜を備えた酵素電極であって、測
定妨害物質除去膜として電解重合膜が用いられているの
で、測定目的物質の測定を正確に行え、そのうえ、製造
も簡単にできる。The enzyme electrode according to the present invention uses hydrogen peroxide as a detection substance and is equipped with a measurement interfering substance removal membrane, and since an electrolytic polymer membrane is used as the measurement interfering substance removal membrane, it is possible to detect the measurement target substance. It allows for accurate measurements and is also easy to manufacture.
代理人 弁理士 松 本 武 彦 ］稿萌俸甫正書（自発 昭和６１年 １月ｌ′７日、 Ｋ ＼。Agent: Patent Attorney Takehiko Matsumoto 】 Written by Mengfufu Seisho (spontaneous) January 1'7, 1985, K＼.
特許庁長官 殿
ｒ−１、事件の表示
ロ訓６０年特許願第２３０４２６号
３、補正をする者
事件との関係 特許出願人
住 所 大阪府門真市大字門真１０４８番
地名 称（５８３）松下電工株式会社
代表者 （ｆｆＥＩｍ役藤 井 貞 夫４、代理人
！
盲
な し
−一６、補正の対象
明細書
７、補正の内容
（１）明細書第３頁第１８行ないし同頁第１９行に「ゲ
ルコールオキシダーゼ」とあるを、「グルコースオキシ
ダーゼ」と訂正する。Commissioner of the Patent Office
r-1, Representation of the case 1960 Patent Application No. 230426 3, Person making the amendment Relationship with the case Patent applicant Address 1048 Kadoma, Kadoma City, Osaka Name (583) Representative of Matsushita Electric Works Co., Ltd. (ffEIm role Sadao Fujii 4, agent! No blindness
-16. Description subject to amendment 7, Contents of amendment (1) From page 3, line 18 of the specification to line 19 of the same page, the term ``gelcol oxidase'' is corrected to ``glucose oxidase.''
Claims (2)
を備えた酵素電極であって、測定妨害物質除去膜として
電解重合膜が用いられていることを特徴とする酵素電極
。(1) An enzyme electrode which uses hydrogen peroxide as a detection substance and is equipped with a measurement interfering substance removal membrane, the enzyme electrode characterized in that an electrolytic polymer membrane is used as the measurement interfering substance removal membrane.
からなる群の中から選ばれた少なくとも１種の電解重合
体である特許請求の範囲第１項記載の酵素電極。(2) The enzyme electrode according to claim 1, wherein the electrolytic polymer membrane is at least one electrolytic polymer selected from the group consisting of pyrrole, thiophene, and aniline.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP60230426A JPS6288953A (en) | 1985-10-15 | 1985-10-15 | Enzyme electrode |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP60230426A JPS6288953A (en) | 1985-10-15 | 1985-10-15 | Enzyme electrode |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPS6288953A true JPS6288953A (en) | 1987-04-23 |
Family
ID=16907707
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP60230426A Pending JPS6288953A (en) | 1985-10-15 | 1985-10-15 | Enzyme electrode |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS6288953A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008256725A (en) * | 2001-05-31 | 2008-10-23 | Instrumentation Lab Co | Analytical instruments and biosensors, and methods for increasing their accuracy and effective life |
| US7632672B2 (en) | 2001-05-31 | 2009-12-15 | Instrumentation Laboratory Co. | Composite membrane containing a cross-linked enzyme matrix for a biosensor |
-
1985
- 1985-10-15 JP JP60230426A patent/JPS6288953A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008256725A (en) * | 2001-05-31 | 2008-10-23 | Instrumentation Lab Co | Analytical instruments and biosensors, and methods for increasing their accuracy and effective life |
| US7632672B2 (en) | 2001-05-31 | 2009-12-15 | Instrumentation Laboratory Co. | Composite membrane containing a cross-linked enzyme matrix for a biosensor |
| 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 |
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