JPH0119880B2 - - Google Patents
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- Publication number
- JPH0119880B2 JPH0119880B2 JP55039726A JP3972680A JPH0119880B2 JP H0119880 B2 JPH0119880 B2 JP H0119880B2 JP 55039726 A JP55039726 A JP 55039726A JP 3972680 A JP3972680 A JP 3972680A JP H0119880 B2 JPH0119880 B2 JP H0119880B2
- Authority
- JP
- Japan
- Prior art keywords
- glucose
- electrode
- enzyme
- immobilized
- concentration
- 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.)
- Expired
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- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 29
- 239000008103 glucose Substances 0.000 claims description 29
- 108010015776 Glucose oxidase Proteins 0.000 claims description 23
- 239000004366 Glucose oxidase Substances 0.000 claims description 23
- 229940116332 glucose oxidase Drugs 0.000 claims description 23
- 235000019420 glucose oxidase Nutrition 0.000 claims description 23
- 102000004190 Enzymes Human genes 0.000 claims description 19
- 108090000790 Enzymes Proteins 0.000 claims description 19
- 229940088598 enzyme Drugs 0.000 claims description 19
- 239000012528 membrane Substances 0.000 claims description 17
- 102000020006 aldose 1-epimerase Human genes 0.000 claims description 14
- 108091022872 aldose 1-epimerase Proteins 0.000 claims description 14
- 108010093096 Immobilized Enzymes Proteins 0.000 claims description 10
- 239000002131 composite material Substances 0.000 claims description 10
- 238000006911 enzymatic reaction Methods 0.000 claims description 4
- 239000000126 substance Substances 0.000 claims description 4
- 150000001875 compounds Chemical class 0.000 claims description 3
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 32
- 230000004044 response Effects 0.000 description 18
- 238000000034 method Methods 0.000 description 8
- 238000006243 chemical reaction Methods 0.000 description 7
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 7
- AZQWKYJCGOJGHM-UHFFFAOYSA-N 1,4-benzoquinone Chemical compound O=C1C=CC(=O)C=C1 AZQWKYJCGOJGHM-UHFFFAOYSA-N 0.000 description 6
- QIGBRXMKCJKVMJ-UHFFFAOYSA-N Hydroquinone Chemical compound OC1=CC=C(O)C=C1 QIGBRXMKCJKVMJ-UHFFFAOYSA-N 0.000 description 6
- 238000005259 measurement Methods 0.000 description 6
- 230000035945 sensitivity Effects 0.000 description 6
- WQZGKKKJIJFFOK-DVKNGEFBSA-N alpha-D-glucose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-DVKNGEFBSA-N 0.000 description 5
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 5
- 230000007423 decrease Effects 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 229910052760 oxygen Inorganic materials 0.000 description 5
- 239000001301 oxygen Substances 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 239000012086 standard solution Substances 0.000 description 5
- 239000004809 Teflon Substances 0.000 description 4
- 229920006362 Teflon® Polymers 0.000 description 4
- 239000007853 buffer solution Substances 0.000 description 4
- 230000003647 oxidation Effects 0.000 description 4
- 238000007254 oxidation reaction Methods 0.000 description 4
- 229910052697 platinum Inorganic materials 0.000 description 4
- 230000009471 action Effects 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- PHOQVHQSTUBQQK-SQOUGZDYSA-N D-glucono-1,5-lactone Chemical compound OC[C@H]1OC(=O)[C@H](O)[C@@H](O)[C@@H]1O PHOQVHQSTUBQQK-SQOUGZDYSA-N 0.000 description 2
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 239000000975 dye Substances 0.000 description 2
- 229960003681 gluconolactone Drugs 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 239000012085 test solution Substances 0.000 description 2
- 230000008901 benefit Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 229920002301 cellulose acetate Polymers 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- ZOMNIUBKTOKEHS-UHFFFAOYSA-L dimercury dichloride Chemical class Cl[Hg][Hg]Cl ZOMNIUBKTOKEHS-UHFFFAOYSA-L 0.000 description 1
- 238000002848 electrochemical method Methods 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- RSAZYXZUJROYKR-UHFFFAOYSA-N indophenol Chemical compound C1=CC(O)=CC=C1N=C1C=CC(=O)C=C1 RSAZYXZUJROYKR-UHFFFAOYSA-N 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 150000003057 platinum Chemical class 0.000 description 1
- -1 potassium ferricyanide Chemical compound 0.000 description 1
- 230000004043 responsiveness Effects 0.000 description 1
- ANRHNWWPFJCPAZ-UHFFFAOYSA-M thionine Chemical compound [Cl-].C1=CC(N)=CC2=[S+]C3=CC(N)=CC=C3N=C21 ANRHNWWPFJCPAZ-UHFFFAOYSA-M 0.000 description 1
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Description
【発明の詳細な説明】
本発明は、固定化されたグルコースオキシダー
ゼと電気化学的電極とを組み合わせてグルコース
濃度を測定する酵素電極の改良に関する。さらに
詳しくは固定化されたグルコースオキシダーゼ
に、さらに固定されたムタロターゼを組み合わせ
た複合固定化酵素を用いることにより、上記複合
酵素固定化反応において生成あるいは消費される
物質濃度を電気化学的に測定しグルコース濃度を
求める方法の測定感度を上げることを目的として
いる。DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an improvement in an enzyme electrode for measuring glucose concentration by combining immobilized glucose oxidase and an electrochemical electrode. More specifically, by using a complex immobilized enzyme that combines immobilized glucose oxidase and further immobilized mutarotase, we can electrochemically measure the concentration of substances produced or consumed in the above complex enzyme immobilization reaction. The purpose is to increase the measurement sensitivity of the method used to determine concentration.
グルコースオキシダーゼと電気化学的電極、例
えば酸素濃度測定用電極あるいは過酸化水素濃度
測定用電極を組み合わせてグルコース濃度を測定
する方法については従来からよく知られている。
一般に、グルコースは、水溶液中では以下に示す
ようにα型とβ型の平衡混合物として存在してい
る。 A method of measuring glucose concentration using a combination of glucose oxidase and an electrochemical electrode, such as an electrode for measuring oxygen concentration or an electrode for measuring hydrogen peroxide concentration, is well known.
Generally, glucose exists in an aqueous solution as an equilibrium mixture of α-type and β-type as shown below.
ここで、α型とβ型の違いは、C1位のOH基の
向きが異なるのみである。現在グルコースオキシ
ダーゼとして知られている酵素は、このα型とβ
型の中でβ型のみに作用し、これをＤ―グルコノ
ラクトンに変換する。 Here, the only difference between the α type and β type is the orientation of the OH group at the C1 position. The enzyme currently known as glucose oxidase is the α-type and β-type.
It acts only on the β type and converts it to D-gluconolactone.
β―Ｄ―グルコース＋O2グルコースオキシダーゼ
――――――――――――→
Ｄ―グルコノラクトン＋H2O2 ……(1)
ここでβ―Ｄ―グルコース濃度が減少するとα
とβ間の平衡値を保つためにα→βの変換反応が
起こりうるが、この反応は上記のグルコースオキ
シダーゼが触媒する反応に比較すると速度が遅
い。したがつてグルコースオキシダーゼ単独酵素
と電気化学的電極を組み合わせてグルコース濃度
を測定する方法では、実際上はαとβの平衡混合
物の中でβ型のみを測定していることになる。 β-D-glucose + O 2 glucose oxidase――――――――――→ D-gluconolactone + H 2 O 2 ...(1) Here, when the β-D-glucose concentration decreases, α
In order to maintain an equilibrium value between and β, an α → β conversion reaction may occur, but this reaction is slow compared to the reaction catalyzed by glucose oxidase described above. Therefore, the method of measuring glucose concentration using a combination of glucose oxidase alone and an electrochemical electrode actually measures only the β form in an equilibrium mixture of α and β.
本発明のグルコース濃度測定用酵素電極では、
α，β間の平衡反応を促進する酵素であるムタロ
ターゼをグルコースオキシダーゼに組み合わせる
ことを特徴としている。この方法によれば、溶液
中のα型グルコースはβ型グルコースにすみやか
に変換されるため、α，β両グルコース濃度を同
時に測定することとなり、結果として測定感度が
向上することになる。以下に本発明の実施例につ
いて述べる。 In the enzyme electrode for measuring glucose concentration of the present invention,
It is characterized by combining mutarotase, an enzyme that promotes the equilibrium reaction between α and β, with glucose oxidase. According to this method, α-type glucose in the solution is quickly converted to β-type glucose, so both α and β glucose concentrations are measured simultaneously, resulting in improved measurement sensitivity. Examples of the present invention will be described below.
実施例 １
グルコースオキシダーゼの凍結乾燥品100mgを
１mlのリン酸緩衝液（PH5.6）中に溶解させ、さ
らにこの混合液中にムタロターゼ6000ユニツト相
当を溶解させる。作製したグルコースオキシダー
ゼとムタロターゼとの混合溶液を市販のガルバニ
ツクセル方式の酵素濃度計に用いるテフロン膜上
に塗布した後、架橋固定化試薬であるグルタルア
ルデヒドを作用させる。このようにしてグルコー
スオキシダーゼームタロターゼ複合酵素固定化テ
フロン膜が作製できる。またムタロターゼを加え
ないて上記と同様にしてグルコースオキシダーゼ
単独固定化テフロン膜を作製した。これら２種類
の膜を上記酸素濃度計の電極先端にとりつけ同一
グルコース標準液に対する応答電流値を比較し
た。Example 1 100 mg of lyophilized glucose oxidase is dissolved in 1 ml of phosphate buffer (PH5.6), and the equivalent of 6000 units of mutarotase is further dissolved in this mixture. The prepared mixed solution of glucose oxidase and mutarotase is applied onto a Teflon membrane used in a commercially available galvanic cell type enzyme concentration meter, and then glutaraldehyde, which is a cross-linking immobilization reagent, is applied. In this way, a Teflon membrane with immobilized glucose oxidase tarotase complex enzyme can be produced. In addition, a Teflon membrane immobilized with glucose oxidase alone was prepared in the same manner as above without adding mutarotase. These two types of membranes were attached to the electrode tip of the oxygen concentration meter, and the response current values to the same glucose standard solution were compared.
第１図は酸素濃度計に接触した緩衝液中にグル
コース標準液を添加し、緩衝液中のグルコース濃
度を０から10-4Mに変化させた場合の酸素濃度計
の応答電流の比較を示した。曲線ａはグルコース
オキシダーゼとムタロターゼとの複合酵素固定化
膜を用いた場合のもので、ｂはグルコースオキシ
ダーゼ単独固定化膜を用いた場合のものである。 Figure 1 shows a comparison of the response current of the oximeter when a glucose standard solution was added to the buffer solution in contact with the oximeter and the glucose concentration in the buffer solution was changed from 0 to 10 -4 M. Ta. Curve a is obtained when a membrane immobilized with a composite enzyme of glucose oxidase and mutarotase is used, and curve b is obtained when a membrane immobilized with glucose oxidase alone is used.
これからわかるように、複合酵素固定化膜を用
いた場合は、単独固定化膜を用いた場合に比較し
て応答電流値の大きさが約1.5倍で応答感度が向
上している。なおガルバニツクセル方式の酸素濃
度計では酸素の還元電流を測定しており、グルコ
ース濃度が上昇すると(1)式に示した反応で酸素濃
度が減少し、結果としてこの還元電流が減少す
る。応答電流の大きさはこの還元電流の減少の大
きさを示している。 As can be seen, when a composite enzyme-immobilized membrane is used, the response current value is approximately 1.5 times greater than when a single immobilized membrane is used, and the response sensitivity is improved. Note that the galvanic cell type oxygen concentration meter measures the reduction current of oxygen, and when the glucose concentration increases, the oxygen concentration decreases due to the reaction shown in equation (1), and as a result, this reduction current decreases. The magnitude of the response current indicates the magnitude of this decrease in reduction current.
実施例 ２
テフロン膜に代えて多孔質セルロースアセテー
ト膜を使用する以外は実施例１と同様にして作製
したグルコースオキシダーゼ単独あるいはムタロ
ターゼとの複合固定化膜を白金電極に密着させ、
この白金電極作用極をポテンシヨスタツトを用い
て、参照電極（飽和カロメル電極）に対し＋
0.6Vの定電位に設定し、対極との間に流れる電
流値を測定する。Example 2 A membrane immobilized with glucose oxidase alone or in combination with mutarotase, prepared in the same manner as in Example 1 except for using a porous cellulose acetate membrane instead of the Teflon membrane, was brought into close contact with a platinum electrode,
Using a potentiostat, this platinum electrode working electrode was
Set to a constant potential of 0.6V, and measure the value of the current flowing between it and the counter electrode.
第２図はこの３電極を組み込んだ測定系の構成
を示す。１は白金電極、１′は白金電極の表面に
保持された固定化酵素膜、２は参照電極、３は対
極、４は緩衝液、５はセパレータである。 FIG. 2 shows the configuration of a measurement system incorporating these three electrodes. 1 is a platinum electrode, 1' is an immobilized enzyme membrane held on the surface of the platinum electrode, 2 is a reference electrode, 3 is a counter electrode, 4 is a buffer solution, and 5 is a separator.
上記の測定系の緩衝液中のグルコース濃度を実
施例１と同様にして０から10-4Mに変化させた場
合の応答電流値を第３図に示した。ｃは複合酵素
固定化膜を用いた場合、ｄは単独酵素固定化膜の
場合である。この結果から複合酵素固定化膜の方
が約1.5倍応答電流が大であり、応答感度の向上
が認められる。なお、ここで測定しているのは前
記反応式(1)によつて生成するH2O2の酸化電流で
ある。 FIG. 3 shows the response current value when the glucose concentration in the buffer solution of the above measurement system was changed from 0 to 10 -4 M in the same manner as in Example 1. c is the case when a composite enzyme-immobilized membrane is used, and d is the case when a single enzyme-immobilized membrane is used. This result shows that the composite enzyme-immobilized membrane has a response current that is about 1.5 times larger and has an improved response sensitivity. Note that what is measured here is the oxidation current of H 2 O 2 generated by the reaction formula (1).
実施例 ３
カーボン電極表面に実施例１で用いたのと同じ
酵素溶液を塗布し、グルタルアルデヒドで処理し
て、電極表面に直接グルコースオキシダーゼ単独
あるいはムタロターゼとの複合酵素固定化膜を形
成する。このようにして作製した２種の電極を実
施例２と同様の測定系に組み入れる。この場合緩
衝液中には10-3Mの濃度でレドツクス化合物の一
種であるＰ―ベンゾキノンが溶解されている。Example 3 The same enzyme solution as used in Example 1 is applied to the surface of a carbon electrode and treated with glutaraldehyde to directly form an enzyme-immobilized membrane containing glucose oxidase alone or mutarotase in combination on the electrode surface. The two types of electrodes produced in this way are incorporated into the same measurement system as in Example 2. In this case, P-benzoquinone, a type of redox compound, is dissolved in the buffer at a concentration of 10 -3 M.
上記酵素固定化カーボン電極を飽和カロメル電
極に対し0.4Vに電位設定し、実施例１，２と同
様グルコース濃度変化（０から１×10-4M）にと
もなう電流値の変化を求めた。 The potential of the enzyme-immobilized carbon electrode was set to 0.4 V with respect to the saturated calomel electrode, and as in Examples 1 and 2, changes in current value with changes in glucose concentration (from 0 to 1×10 −4 M) were determined.
第４図に複合酵素固定化の場合ｅと単独酵素固
定化の場合ｆとの比較を示す。これから複合固定
化酵素を用いる場合の方が約1.5倍応答感度が大
であることがわかる。ここでは、式(1)のO2にか
わつてＰ―ベンゾキノンが酵素反応にともなつて
ヒドロキノンになり、この生成したヒドロキノン
の酸化電流を測定していることになる。 FIG. 4 shows a comparison between case e of composite enzyme immobilization and case f of single enzyme immobilization. This shows that the response sensitivity is about 1.5 times higher when using a complex immobilized enzyme. Here, instead of O 2 in formula (1), P-benzoquinone becomes hydroquinone through an enzymatic reaction, and the oxidation current of the generated hydroquinone is measured.
レドツクス化合物としてはＰ―ベンゾキノン以
外にチオニン、インドフエノール、フエリシアン
化カリ等のレドツクス色素を用いることができ
る。またこれらのレドツクス色素はすでに本発明
者らが提案しているごとく電極に固定化して用い
ることも可能である。 As the redox compound, in addition to P-benzoquinone, redox dyes such as thionine, indophenol, and potassium ferricyanide can be used. Furthermore, these redox dyes can also be used by being immobilized on electrodes as already proposed by the present inventors.
さらにグルコースオキシダーゼとムタロターゼ
の固定化の割合は、ムタロターゼの活性がグルコ
ースオキシダーゼの活性に比較して十分高いこと
が望ましい。 Furthermore, regarding the immobilization ratio of glucose oxidase and mutarotase, it is desirable that the activity of mutarotase is sufficiently higher than that of glucose oxidase.
以上実施例１〜３いずれにおいてもムタロター
ゼとの複合固定化酵素を用いる方法がグルコース
オキシダーゼ単独を用いる方法に比較して約1.5
倍の応答電流値を示している。水溶液中（20℃）
においてグルコースのα型とβ型の比率α／βは
36／64であることが知られており、この数字を考
慮すると上記の1.5倍という数字はほぼ説明がつ
く。すなわちαグルコースがムタロターゼの作用
できわめてすみやかにβグルコースに変換される
とともに、グルコースオキシダーゼの作用を受け
るために、結果として溶液中のαグルコースもβ
グルコースもともにグルコースオキシダーゼの作
用を受けることになり、本発明のグルコース濃度
測定用酵素電極を用いた方法ではαグルコースと
βグルコースの総グルコース濃度を測定している
わけである。実際α型も同時に反応に関与すると
すると36＋64／64＝1.56倍の応答が得られることに
なり、この値は実施例で得られている値にかなり
近い。 In all of Examples 1 to 3 above, the method using a complex immobilized enzyme with mutarotase was approximately 1.5% higher than the method using glucose oxidase alone.
The response current value is twice as large. In aqueous solution (20℃)
The ratio α/β of the α-form and β-form of glucose is
It is known that the ratio is 36/64, and considering this number, the above number of 1.5 times can be explained. In other words, α-glucose is very quickly converted to β-glucose by the action of mutarotase, and since it is also subjected to the action of glucose oxidase, as a result, α-glucose in the solution is also converted to β-glucose.
Both glucose is affected by the action of glucose oxidase, and the method using the enzyme electrode for measuring glucose concentration of the present invention measures the total glucose concentration of α-glucose and β-glucose. In fact, if the α type also participates in the reaction at the same time, a response that is 36+64/64=1.56 times greater will be obtained, and this value is quite close to the value obtained in the example.
グルコースオキシダーゼ単独を電極に固定化し
た酵素電極によつてグルコース全濃度を測定する
には、濃度測定を目的とするグルコース含有被検
液の応答と濃度既知の標準液の濃度とを比較する
ことによつて行われる。すなわち濃度測定を目的
とするグルコース含有液と濃度既知のグルコース
標準液とでαとβの含有割合が一定であると仮定
して両液のβ型に対して得られた応答を比較して
求めているわけである。ところが実際の被検液で
は液の温度、PH、夾雑物等によつてαとβの平衡
値は当然変動しうる。そして標準液を被検液とま
つたく同じα，β間干衡状態に保つことは当然の
ことながら困難であり、分析の誤差をまねきやす
い。このことからα，β両グルコースに活性を有
する複合酵素系を用いる本発明の方法は、より精
度良く全グルコース濃度を測定しうるという利点
も有している。 To measure the total glucose concentration using an enzyme electrode in which glucose oxidase alone is immobilized on the electrode, it is necessary to compare the response of a glucose-containing test solution whose concentration is to be measured with the concentration of a standard solution whose concentration is known. It is done by folding. In other words, assuming that the content ratio of α and β is constant between a glucose-containing solution whose concentration is to be measured and a glucose standard solution of known concentration, the response obtained for the β-type of both solutions is compared. That's why. However, in an actual test liquid, the equilibrium values of α and β may vary depending on the temperature, pH, impurities, etc. of the liquid. Naturally, it is difficult to maintain the standard solution in the same equilibrium state between α and β as the test solution, which tends to lead to analytical errors. Therefore, the method of the present invention using a composite enzyme system having activity on both α and β glucose also has the advantage of being able to measure the total glucose concentration with higher accuracy.
また、上記２種の酵素をリン酸緩衝液に溶解し
た状態で室温保存すると急速に活性が低下するの
に対して、実施例１〜３に示した複合固定化酵素
系においては、緩衝液中300日の室温保存後にお
いても初期の約75％の応答性を維持しており、保
存性に優れたものであつた。 Furthermore, if the above two types of enzymes are dissolved in phosphate buffer and stored at room temperature, the activity rapidly decreases, whereas in the composite immobilized enzyme system shown in Examples 1 to 3, the activity is Even after storage at room temperature for 300 days, it maintained approximately 75% of its initial responsiveness, indicating excellent storage stability.
以上のようにムタロターゼとグルコースオキシ
ダーゼからなる複合固定化酵素を用い、グルコー
ス濃度を電気化学的に測定する酵素電極において
は、全グルコース濃度を測定していることにな
り、応答感度、測定精度が向上し、さらには保存
性にも優れるという効果がある。 As described above, the enzyme electrode that electrochemically measures glucose concentration using a composite immobilized enzyme consisting of mutarotase and glucose oxidase measures the total glucose concentration, improving response sensitivity and measurement accuracy. Furthermore, it has the effect of being excellent in storage stability.
第１図は酸素濃度計の応答曲線を示す図、第２
図は３電極電気化学測定系の構成図、第３図は
H2O2の酸化電流を測定する場合の電流応答曲線、
第４図はヒドロキノンの酸化電流を測定する場合
の電流応答曲線である。
Figure 1 shows the response curve of the oximeter, Figure 2 shows the response curve of the oximeter.
The figure is a configuration diagram of a three-electrode electrochemical measurement system, and Figure 3 is
Current response curve when measuring the oxidation current of H 2 O 2 ,
FIG. 4 is a current response curve when measuring the oxidation current of hydroquinone.
Claims (1)
ロターゼからなる複合固定化酵素膜を電極近傍に
備えてなり、複合固定化酵素反応により生成ある
いは消費される物質の濃度を前記電極で電気化学
的に検知することを特徴とするグルコース濃度測
定用酵素電極。 ２ 複合固定化酵素反応により生成される物質
が、H2O2あるいは還元型のレドツクス化合物で
ある特許請求の範囲第１項に記載のグルコース濃
度測定用酵素電極。 ３ 複合固定化酵素反応により消費される物質
が、O2である特許請求の範囲第１項に記載のグ
ルコース濃度測定用酵素電極。[Scope of Claims] 1. A composite immobilized enzyme membrane consisting of immobilized glucose oxidase and mutarotase is provided near an electrode, and the concentration of a substance produced or consumed by the composite immobilized enzyme reaction is measured electrochemically at the electrode. An enzyme electrode for measuring glucose concentration, which is characterized by its ability to detect glucose concentration. 2. The enzyme electrode for measuring glucose concentration according to claim 1, wherein the substance produced by the complex immobilized enzyme reaction is H 2 O 2 or a reduced redox compound. 3. The enzyme electrode for measuring glucose concentration according to claim 1, wherein the substance consumed by the complex immobilized enzyme reaction is O2 .
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP3972680A JPS56137899A (en) | 1980-03-27 | 1980-03-27 | Determining method of glucose concentration |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP3972680A JPS56137899A (en) | 1980-03-27 | 1980-03-27 | Determining method of glucose concentration |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS56137899A JPS56137899A (en) | 1981-10-28 |
| JPH0119880B2 true JPH0119880B2 (en) | 1989-04-13 |
Family
ID=12560978
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP3972680A Granted JPS56137899A (en) | 1980-03-27 | 1980-03-27 | Determining method of glucose concentration |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS56137899A (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6024444A (en) * | 1983-07-19 | 1985-02-07 | Matsushita Electric Ind Co Ltd | Bio-sensor |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS4973193A (en) * | 1972-09-28 | 1974-07-15 | ||
| JPS5236092A (en) * | 1975-09-17 | 1977-03-19 | Omron Tateisi Electronics Co | Measuring electrode |
| JPS5258995A (en) * | 1975-11-07 | 1977-05-14 | Owens Illinois Inc | Measuring method and apparatus for glucose |
-
1980
- 1980-03-27 JP JP3972680A patent/JPS56137899A/en active Granted
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS4973193A (en) * | 1972-09-28 | 1974-07-15 | ||
| JPS5236092A (en) * | 1975-09-17 | 1977-03-19 | Omron Tateisi Electronics Co | Measuring electrode |
| JPS5258995A (en) * | 1975-11-07 | 1977-05-14 | Owens Illinois Inc | Measuring method and apparatus for glucose |
Also Published As
| Publication number | Publication date |
|---|---|
| JPS56137899A (en) | 1981-10-28 |
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