JPH0332742B2 - - Google Patents
Info
- Publication number
- JPH0332742B2 JPH0332742B2 JP57100213A JP10021382A JPH0332742B2 JP H0332742 B2 JPH0332742 B2 JP H0332742B2 JP 57100213 A JP57100213 A JP 57100213A JP 10021382 A JP10021382 A JP 10021382A JP H0332742 B2 JPH0332742 B2 JP H0332742B2
- Authority
- JP
- Japan
- Prior art keywords
- electrode
- glucose
- sucrose
- mutarotase
- enzyme
- 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 - Lifetime
Links
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 claims description 36
- 229930006000 Sucrose Natural products 0.000 claims description 26
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 26
- 239000005720 sucrose Substances 0.000 claims description 26
- 229910052697 platinum Inorganic materials 0.000 claims description 18
- 102000004190 Enzymes Human genes 0.000 claims description 17
- 108090000790 Enzymes Proteins 0.000 claims description 17
- 229940088598 enzyme Drugs 0.000 claims description 17
- 239000012528 membrane Substances 0.000 claims description 15
- 102000020006 aldose 1-epimerase Human genes 0.000 claims description 14
- 108091022872 aldose 1-epimerase Proteins 0.000 claims description 14
- 108010015776 Glucose oxidase Proteins 0.000 claims description 13
- 239000004366 Glucose oxidase Substances 0.000 claims description 13
- 229940116332 glucose oxidase Drugs 0.000 claims description 13
- 235000019420 glucose oxidase Nutrition 0.000 claims description 13
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims description 11
- 239000007788 liquid Substances 0.000 claims description 8
- 238000012360 testing method Methods 0.000 claims description 8
- 108010051210 beta-Fructofuranosidase Proteins 0.000 claims description 6
- 235000011073 invertase Nutrition 0.000 claims description 6
- 239000000126 substance Substances 0.000 claims description 5
- 238000001514 detection method Methods 0.000 claims description 4
- 230000001590 oxidative effect Effects 0.000 claims description 2
- 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 20
- 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 description 18
- 239000008103 glucose Substances 0.000 description 13
- 238000000034 method Methods 0.000 description 9
- 239000011148 porous material Substances 0.000 description 7
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 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
- 238000005259 measurement Methods 0.000 description 5
- 230000003647 oxidation Effects 0.000 description 5
- 238000007254 oxidation reaction Methods 0.000 description 5
- 229910021607 Silver chloride Inorganic materials 0.000 description 4
- HKZLPVFGJNLROG-UHFFFAOYSA-M silver monochloride Chemical compound [Cl-].[Ag+] HKZLPVFGJNLROG-UHFFFAOYSA-M 0.000 description 4
- 239000012085 test solution Substances 0.000 description 4
- 235000010323 ascorbic acid Nutrition 0.000 description 3
- 229960005070 ascorbic acid Drugs 0.000 description 3
- 239000011668 ascorbic acid Substances 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 230000002452 interceptive effect Effects 0.000 description 3
- 238000000691 measurement method Methods 0.000 description 3
- 238000004544 sputter deposition Methods 0.000 description 3
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 2
- 238000006911 enzymatic reaction Methods 0.000 description 2
- 239000008055 phosphate buffer solution Substances 0.000 description 2
- RFSUNEUAIZKAJO-VRPWFDPXSA-N D-Fructose Natural products OC[C@H]1OC(O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-VRPWFDPXSA-N 0.000 description 1
- RGHNJXZEOKUKBD-UHFFFAOYSA-N D-gluconic acid Natural products OCC(O)C(O)C(O)C(O)C(O)=O RGHNJXZEOKUKBD-UHFFFAOYSA-N 0.000 description 1
- RGHNJXZEOKUKBD-SQOUGZDYSA-N Gluconic acid Natural products OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C(O)=O RGHNJXZEOKUKBD-SQOUGZDYSA-N 0.000 description 1
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 description 1
- 108010093096 Immobilized Enzymes Proteins 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005868 electrolysis reaction Methods 0.000 description 1
- 239000010408 film Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000007429 general method Methods 0.000 description 1
- 235000012208 gluconic acid Nutrition 0.000 description 1
- 239000000174 gluconic acid Substances 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229920000515 polycarbonate Polymers 0.000 description 1
- 239000004417 polycarbonate Substances 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 239000010409 thin film Substances 0.000 description 1
- 238000007740 vapor deposition Methods 0.000 description 1
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
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Analytical Chemistry (AREA)
- Immunology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Description
【発明の詳細な説明】
本発明は、被検液中のスクロース（しよ糖）の
濃度を精度よく、しかも迅速簡便に測定すること
のできるスクロース濃度測定用酵素電極に関す
る。DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an enzyme electrode for measuring sucrose concentration that can accurately, quickly and easily measure the concentration of sucrose in a test liquid.
スクロースは、食品中に広く含有される糖の１
つとして重要である。従来、スクロース濃度の測
定に関しては、化学反応を利用する方法、あるい
は液体クロマトグラフイーなど種々の測定法が用
いられているが、精度や迅速性に欠けたり、ある
いは測定操作が煩雑であるなどの問題点を有する
ものである。これに対し酵素反応を利用する測定
法、特に酵素を電極近傍に固定化したいわゆる酵
素電極を用いる方法は、上記欠点を解決すること
のできる測定法として注目されている。酵素法に
よるスクロースの測定方法について以下に反応式
で示す。 Sucrose is one of the sugars widely found in foods.
It is important as one of the Conventionally, various measurement methods have been used to measure sucrose concentration, such as methods using chemical reactions or liquid chromatography, but these methods lack accuracy and speed, or have complicated measurement operations. This has some problems. On the other hand, a measurement method using an enzyme reaction, particularly a method using a so-called enzyme electrode in which an enzyme is immobilized near the electrode, is attracting attention as a measurement method that can solve the above-mentioned drawbacks. The method for measuring sucrose using an enzymatic method is shown below using a reaction formula.
スクロース＋H2Oβ−フルクトシダーゼ
――――――――――――→
α−Ｄ−グルコース＋Ｄ−フルクトース (1)
α−Ｄ−グルコースムタロターゼ
―――――――→
β−Ｄ−グルコース (2)
β−Ｄ−グルコース＋H2O＋O2
グルコースオキシダーゼ
―――――――――――――→
H2O2＋グルコン酸 (3)
H2O2→2H＋O2＋2e (4)
まず、被検液中のスクロースが(1)式の様にβ−
フルクトシダーゼによりα−Ｄ−グルコースに分
解され、続いて(2)式のようにムタロターゼの作用
によりβ−Ｄ−グルコースとなり、さらに(3)式の
様にグルコースオキシダーゼにより過酸化水素
（H2O2）を生成する。そこで、このH2O2を(4)式
のように白金電極などからなる過酸化水素検出用
の電極を用いて電解酸化することにより、得られ
る酸化電流値からもとの被検液中に含まれるスク
ロース濃度を知ることができる。Sucrose + H 2 O β-fructosidase――――――――――――→ α-D-glucose + D-fructose (1) α-D-glucose mutarotase――――――――→ β- D-glucose (2) β-D-glucose + H 2 O + O 2 glucose oxidase――――――――――――→ H 2 O 2 + gluconic acid (3) H 2 O 2 →2H + O 2 +2e ( 4) First, the sucrose in the test solution is β- as shown in equation (1).
It is decomposed into α-D-glucose by fructosidase, then becomes β-D-glucose by the action of mutarotase as shown in equation (2), and then hydrogen peroxide (H 2 O 2 ) is generated. Therefore, by electrolytically oxidizing this H 2 O 2 using an electrode for detecting hydrogen peroxide, such as a platinum electrode, as shown in equation (4), the oxidation current value obtained indicates that You can know the concentration of sucrose contained in it.
しかし、この方法においては、スクロースをグ
ルコースに変換することに基づいているため、被
検液中にグルコースが共存する場合には誤差を生
じることになる。この対策としては、従来の一般
的方法として、２つの電極を用いる方法が考えら
れる。すなわち、β−フルクトシダーゼとムタロ
ターゼおよびグルコースオキシダーゼの３種類の
酵素を固定化したスクロース検出用（共存グルコ
ースも同時に検出する）の電極と、ムタロターゼ
とグルコースオキシダーゼの２種類の酵素を固定
化した共存グルコース検出用の電極を使用し、こ
の２つの電極で得られる電流値を差し引くことに
より被検液中のスクロースに基づく電流値のみを
得るという方法である。しかし、この方法は、電
極を２つ用いることの他に、２つの電極の応答特
性をうまくバランスさせる必要があるなど、実用
的には甚だ不便である。 However, since this method is based on converting sucrose to glucose, errors will occur if glucose coexists in the test liquid. As a countermeasure against this problem, a conventional and general method using two electrodes can be considered. In other words, an electrode for sucrose detection (detects coexisting glucose at the same time) on which three types of enzymes, β-fructosidase, mutarotase, and glucose oxidase are immobilized, and a coexistence electrode on which two types of enzymes, mutarotase and glucose oxidase, are immobilized. This method uses an electrode for glucose detection and subtracts the current values obtained from these two electrodes to obtain only the current value based on sucrose in the test liquid. However, in addition to using two electrodes, this method requires a good balance between the response characteristics of the two electrodes, which is extremely inconvenient from a practical standpoint.
そこで、本発明は、以上に述べた不都合をなく
し、迅速かつ容易に、繰り返し使用でき、しかも
共存グルコースの影響を除去することのできるス
クロース濃度測定用酵素電極を提供するものであ
る。 Therefore, the present invention provides an enzyme electrode for measuring sucrose concentration that eliminates the above-mentioned disadvantages, can be used quickly, easily, and repeatedly, and can eliminate the influence of coexisting glucose.
本発明のスクロース濃度測定用酵素電極の特徴
は、グルコースオキシダーゼとムタロターゼおよ
びβ−フルクトシダーゼを固定化した過酸化水素
検出用の第１の電極と、第１の電極による過酸化
水素の検出を妨害する物質を電解除去するための
第２の電極とからなり、第２の電極上にグルコー
スオキシダーゼとムタロターゼを固定化した点に
ある。ここで、第１の電極および第２の電極は、
それぞれ白金層を形成した多孔質膜から構成する
のがよく、第２の電極は、第１の電極に対して被
検液側に配置されている。 The enzyme electrode for measuring sucrose concentration of the present invention is characterized by a first electrode for detecting hydrogen peroxide on which glucose oxidase, mutarotase, and β-fructosidase are immobilized, and a detection of hydrogen peroxide by the first electrode. It consists of a second electrode for electrolytically removing interfering substances, and glucose oxidase and mutarotase are immobilized on the second electrode. Here, the first electrode and the second electrode are
It is preferable that each electrode is composed of a porous membrane on which a platinum layer is formed, and the second electrode is arranged on the side of the test liquid with respect to the first electrode.
第１図は本発明の酵素電極の一実施例について
部分拡大断面模式図で示す。図中、１は第１の電
極、２は第の電極である。第１の電極は、多孔質
膜３の片側の膜面上にスパツタリング、蒸着など
の方法で白金層４を形成し、さらに膜面上および
孔中にグルコースオキシダーゼとムタロターゼお
よびβ−フルクトシダーゼを固定化している。５
はこれらの酵素固定化層である。第２の電極２
は、多孔質膜６の両面に白金層７を前記同様の方
法により形成し、孔中および膜面上にはグルコー
スオキシダーゼとムタロターゼを固定化してい
る。８は酵素固定化層である。さらに、これら２
つの電極は一体化され、一枚の薄膜状の酵素電極
に構成されている。測定に際しては、第２の電極
側を被検液側となる様にして用いる。 FIG. 1 is a partially enlarged schematic cross-sectional view of an embodiment of the enzyme electrode of the present invention. In the figure, 1 is a first electrode, and 2 is a second electrode. In the first electrode, a platinum layer 4 is formed on one side of the porous membrane 3 by sputtering, vapor deposition, etc., and glucose oxidase, mutarotase, and β-fructosidase are added on the membrane surface and in the pores. It is fixed. 5
are these enzyme immobilization layers. second electrode 2
In this method, platinum layers 7 are formed on both sides of a porous membrane 6 by a method similar to that described above, and glucose oxidase and mutarotase are immobilized in the pores and on the membrane surface. 8 is an enzyme immobilization layer. Furthermore, these two
The two electrodes are integrated into a single thin-film enzyme electrode. During measurement, the second electrode is used with the second electrode facing the test liquid side.
上記の酵素電極によるスクロース濃度測定につ
いて、前述の(1)〜(4)の反応式に基づいて説明す
る。すなわち、被検液中のスクロースは第２の電
極の孔中を通つて第１の電極に拡散し、β−フル
クトシダーゼの作用でα−Ｄ−グルコースとな
り、次にムタロターゼによつてβ−Ｄ−グルコー
スとなり、さらにグルコースオキシダーゼによつ
てH2O2が生成する。第１の電極の電位をH2O2の
十分な酸化電位に設定しておくことにより、
H2O2が白金層で酸化される。このとき得られる
酸化電流は被検液中のスクロース濃度に依存す
る。 The measurement of sucrose concentration using the enzyme electrode described above will be explained based on the reaction formulas (1) to (4) described above. That is, sucrose in the test solution diffuses into the first electrode through the pores of the second electrode, becomes α-D-glucose by the action of β-fructosidase, and then becomes β-D-glucose by mutarotase. D-glucose is formed, and H 2 O 2 is further generated by glucose oxidase. By setting the potential of the first electrode to a sufficient oxidation potential of H 2 O 2 ,
H 2 O 2 is oxidized in the platinum layer. The oxidation current obtained at this time depends on the sucrose concentration in the test liquid.
被検液中にα又はβのＤ−グルコースが共存す
る場合には、第２の電極に固定化されたグルコー
スオキシダーゼとムタロターゼの作用により(2)、
(3)式と同様にH2O2が生成し、ただちに白金層７
によつて(4)式のように電解酸化される。また、ア
スコルビン酸をはじめとする白金層で直接電解さ
れやすい妨害物質が共存する場合にも、白金層７
で電解酸化される。すなわち、第２の電極で前も
つて電解酸化することにより、第１の電極におけ
るスクロースのみに基づいて生成するH2O2の電
解酸化に対する妨害物質の影響を除去することが
できる。 When α or β D-glucose coexists in the test solution, due to the action of glucose oxidase and mutarotase immobilized on the second electrode (2),
Similar to equation (3), H 2 O 2 is generated and immediately the platinum layer 7
is electrolytically oxidized as shown in equation (4). In addition, when interfering substances such as ascorbic acid that are easily electrolyzed directly in the platinum layer coexist, the platinum layer 7
is electrolytically oxidized. That is, by performing electrolytic oxidation beforehand at the second electrode, it is possible to eliminate the influence of interfering substances on the electrolytic oxidation of H 2 O 2 produced only based on sucrose at the first electrode.
以上のように、白金層を形成した多孔質膜にム
タロターゼとグルコースオキシダーゼを固定化し
た第２の電極を用いることにより、共存するグル
コースやアスコルビン酸など、第１の電極に対し
て妨害となる物質の影響を容易に除去することが
できる。 As described above, by using a second electrode in which mutarotase and glucose oxidase are immobilized on a porous membrane formed with a platinum layer, substances that interfere with the first electrode, such as coexisting glucose and ascorbic acid, can be removed. The influence of can be easily removed.
以下、本発明の一実施例を説明する。 An embodiment of the present invention will be described below.
孔径2000Å、孔密度３×108個／cm2、膜厚10μｍ
のポリカーボネート多孔質膜の片面に厚さが約
1000Åの白金層をスパツタリングにより形成し、
次にこの白金層の側からグルコースオキシダーゼ
とムタロターゼおよびβ−フルクトシダーゼの混
合液を膜面上および孔中に展開した後、乾燥し、
グルタルアルデヒド蒸気中において、25℃で60分
間架橋固定化を行い、この後、十分洗浄し、第１
の電極とした。次に、第２の電極としては、上記
と同じ多孔質膜を使用し、膜の両面にスパツタリ
ングにより上記同様の白金層を形成し、次にグル
コースオキシダーゼとムタロターゼの混合液を膜
の両面および孔中に展開、乾燥後、前記同様に固
定化した。 Pore diameter 2000Å, pore density 3× 108 / cm2 , film thickness 10μm
One side of the polycarbonate porous membrane has a thickness of approx.
A 1000 Å platinum layer was formed by sputtering,
Next, a mixture of glucose oxidase, mutarotase, and β-fructosidase is spread on the membrane surface and into the pores from the platinum layer side, and then dried.
Cross-linking fixation was carried out in glutaraldehyde vapor at 25°C for 60 minutes, after which the first
It was used as an electrode. Next, as the second electrode, the same porous membrane as above is used, a platinum layer similar to the above is formed by sputtering on both sides of the membrane, and then a mixed solution of glucose oxidase and mutarotase is applied to both sides of the membrane and the pores. After spreading and drying, it was fixed in the same manner as above.
得られた第１の電極と第２の電極を第１図に示
した様に重ね合わせた後、第２図に示す電極ホル
ダーに組み込んで試験に供した。 After the obtained first electrode and second electrode were overlapped as shown in FIG. 1, they were assembled into an electrode holder shown in FIG. 2 and subjected to a test.
第２図において、９は上記のスクロース濃度測
定用酵素電極であり、円筒形の樹脂製ホルダー１
０，１１によつて固定され、第１の電極はリード
１２に、第２の電極はリード１３にそれぞれ接続
されている。ホルダー内部には、第１の電極に対
する白金対極１４とAg／AgCl参照極１５を備
え、PH5.6のリン酸緩衝液１６が満たされている。
またホルダー外部には、第２の電極に対する白金
対極１７とAg／AgCl参照極１８を備えている。
また、第１および第２の電極の位置関係について
は、ホルダー内側に第１の電極が、ホルダー外側
（被検液側）に第２の電極がそれぞれ位置する様
に装着されている。 In FIG. 2, 9 is the enzyme electrode for measuring sucrose concentration, and the cylindrical resin holder 1
0 and 11, the first electrode is connected to the lead 12, and the second electrode is connected to the lead 13, respectively. The inside of the holder includes a platinum counter electrode 14 and an Ag/AgCl reference electrode 15 for the first electrode, and is filled with a phosphate buffer solution 16 of pH 5.6.
Furthermore, a platinum counter electrode 17 and an Ag/AgCl reference electrode 18 for the second electrode are provided outside the holder.
Regarding the positional relationship between the first and second electrodes, the first electrode is installed inside the holder, and the second electrode is installed outside the holder (on the side of the sample liquid).
上記の電極系をPH5.6のリン酸緩衝液中に浸漬
し、第１の電極および第２の電極の電位をそれぞ
れ＋0.60V vs.Ag／AgClに設定した。次にスク
ロース含む被検液を添加したところ、第１の電極
の電流は急激に増加し、約１分程度で定常値に達
した。このときの電流増加量とスクロース濃度の
関係を第３図にＡで示す。同様にして、グルコー
スを添加した場合のグルコース濃度と電流増加量
の関係をＢで示す。 The above electrode system was immersed in a phosphate buffer solution with a pH of 5.6, and the potentials of the first electrode and the second electrode were each set to +0.60 V vs.Ag/AgCl. Next, when a test solution containing sucrose was added, the current at the first electrode rapidly increased and reached a steady value in about 1 minute. The relationship between the amount of current increase and the sucrose concentration at this time is shown by A in FIG. Similarly, B indicates the relationship between the glucose concentration and the amount of increase in current when glucose is added.
比較のために、第２の電極に全く電位を印加せ
ず、第１の電極だけ＋0.60V vs.Ag／AgClに設
定しておき、上記同様にグルコースに対する電流
増加量を測定した。これをＣで示す。 For comparison, no potential was applied to the second electrode, only the first electrode was set at +0.60V vs.Ag/AgCl, and the amount of increase in current relative to glucose was measured in the same manner as above. This is denoted by C.
第３図のＢとＣを比較すれば明らかなように、
第２の電極で電解を行うことにより、グルコース
に基づく電流増加を大幅に減少させることができ
る。また、スクロースに対しては、Ａに示すよう
に良好な直線性を有するものであつた。また、こ
れとは別にアスコルビン酸に対して測定したとこ
ろ、上記グルコースの場合と同様にほとんど影響
を受けないことがわかつた。 As is clear from comparing B and C in Figure 3,
By performing electrolysis at the second electrode, the glucose-based current increase can be significantly reduced. Moreover, as shown in A, it had good linearity with respect to sucrose. Separately, ascorbic acid was measured and found to have almost no effect, similar to the case with glucose.
さらに、上記測定を長期間にわたつて繰り返し
行つたところ、酵素電極の応答特性はほとんど低
下することなく、安定であつた。 Furthermore, when the above measurements were repeated over a long period of time, the response characteristics of the enzyme electrode remained stable with almost no deterioration.
本発明のスクロース濃度測定用酵素電極に関し
て、多孔質膜の種類、白金層の形状、２つの電極
の構成、さらには設定電位をはじめとする測定条
件などについては、実施例に述べたものに限定さ
れることはなく、本発明の主旨に沿つたものであ
れば良い。 Regarding the enzyme electrode for measuring sucrose concentration of the present invention, measurement conditions such as the type of porous membrane, the shape of the platinum layer, the configuration of the two electrodes, and the set potential are limited to those described in the examples. It is not necessary to do so as long as it is in line with the gist of the present invention.
第１図は本発明によるスクロース濃度測定用酵
素電極の一実施例を示す部分拡大断面模式図、第
２図はスクロース濃度を測定するための電極ホル
ダーと電極系を示す模式図、第３図はスクロース
濃度あるいはグルコース濃度と第１の電極の電流
増加量の関係を示す図である。
１……第１の電極、２……第２の電極、３，６
……多孔質膜、４，７……白金層、５，８……固
定化酵素層。
FIG. 1 is a schematic partially enlarged cross-sectional view showing an embodiment of the enzyme electrode for measuring sucrose concentration according to the present invention, FIG. 2 is a schematic view showing an electrode holder and electrode system for measuring sucrose concentration, and FIG. FIG. 3 is a diagram showing the relationship between sucrose concentration or glucose concentration and the amount of increase in current of the first electrode. 1...First electrode, 2...Second electrode, 3,6
...Porous membrane, 4,7...Platinum layer, 5,8...Immobilized enzyme layer.
Claims (1)
極による過酸化水素の検出を妨害する物質を電解
酸化するための第２の電極とを有し、第１の電極
および第２の電極はそれぞれ白金層を形成した多
孔質膜からなり、第１の電極にグルコースオキシ
ダーゼとムタロターゼおよびβ−フルクトシダー
ゼを固定化し、第２の電極にグルコースオキシダ
ーゼとムタロターゼを固定化してなり、第２の電
極を第１の電極に対して被検液側に配置したこと
を特徴とするスクロース濃度測定用酵素電極。1. A first electrode for detecting hydrogen peroxide, and a second electrode for electrolytically oxidizing a substance that interferes with detection of hydrogen peroxide by the first electrode. The electrodes each consist of a porous membrane on which a platinum layer is formed, and glucose oxidase, mutarotase, and β-fructosidase are immobilized on the first electrode, glucose oxidase and mutarotase are immobilized on the second electrode, and the second electrode is made of a porous membrane with a platinum layer formed thereon. An enzyme electrode for measuring sucrose concentration, characterized in that an electrode is disposed on the test liquid side with respect to the first electrode.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP57100213A JPS58216947A (en) | 1982-06-10 | 1982-06-10 | Enzyme electrode for measuring sucrose concentration |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP57100213A JPS58216947A (en) | 1982-06-10 | 1982-06-10 | Enzyme electrode for measuring sucrose concentration |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS58216947A JPS58216947A (en) | 1983-12-16 |
| JPH0332742B2 true JPH0332742B2 (en) | 1991-05-14 |
Family
ID=14268018
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP57100213A Granted JPS58216947A (en) | 1982-06-10 | 1982-06-10 | Enzyme electrode for measuring sucrose concentration |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS58216947A (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6193948A (en) * | 1984-10-13 | 1986-05-12 | Shimadzu Corp | Sucrose sensor |
| US5225321A (en) * | 1987-09-11 | 1993-07-06 | Kanzaki Paper Mfg., Co., Ltd. | Measuring apparatus using enzyme electrodes and the method thereof |
| US5206145A (en) * | 1988-05-19 | 1993-04-27 | Thorn Emi Plc | Method of measuring the concentration of a substance in a sample solution |
| KR100481663B1 (en) * | 2002-09-24 | 2005-04-08 | 김희찬 | Biosensor contained mesoporous platinum and method of determining concentration using same |
-
1982
- 1982-06-10 JP JP57100213A patent/JPS58216947A/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| JPS58216947A (en) | 1983-12-16 |
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