JP2004251913A - 電気化学センサ - Google Patents
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Abstract
【構成】 (a) 白金化炭素粉末粒子を中性緩衝液と混合し、
(b) 上記工程(a) で得た緩衝化した白金化炭素粉末粒子をタンパク質と混合してタンパク質−白金化活性炭組成物を形成し、
(c) 上記工程(b) で得た前記タンパク質−白金化活性炭組成物を結合剤樹脂と混合し、
(d) 上記工程(c) で得た混合物を混練し、
(e) 上記工程(d) で得た混練生成物に酵素または不活性タンパク質のいずれかを加えて混合してペーストを形成する。
【選択図】 図15
Description
C6H12O6+O2+H2O−−−−−−−>C6H12O7+H2O2
H2O2 −−−−−−−>O2+2H++2e−
これらの反応において、グルコースは酸素と反応してグルコノラクトンおよび過酸化水素を生成する。そこで、適切な電極を用いて、電気信号として過酸化水素の生成を測定することができる。この信号は、過酸化物からの電子が電極に移送された後に生成され、適切な条件下で、酵素を触媒とする電流がグルコースの濃度に比例する。酵素電極等のラクテート電極センサも同様に、ラクテートオキシダーゼのような酵素の存在下でラクテートを転化する。
極性基体の両方に対して優れた接着特性を有することである。
規定される電極の縁による表面積を考慮していない。このような縁の寸法は、電極の全体の面積と比較して小さい。したがって、露出した表面積の規格値はいくぶん近似値である。
図13、14および32並びに表Iを参照する。実質的に本発明の実施態様による構成部材および設計による固体プレーナグルコースセンサ28のある適切な構成を以下の要素の組合せにより提供する。
作用電極90に使用するために活性層96を調製した(実施例I参照)。グルコースセンサ用の活性層96は主に、イーテック社から得られる白金化炭素粉末粒子に固定化された、バイオザイムラボラトリーズ社から得られるグルコースオキシダーゼを触媒的に活性である量含み、これらの粒子は層中に実質的に均一に分布している。
濾過し、100 mlの緩衝液を使用して減圧濾過を行ないながら中和した炭素を洗浄した。緩衝液の大部分を炭素を通して引いた後に約15から20秒間に亘り減圧状態を維持して、炭素をやや乾燥させて、材料の取扱い適性を改良した。
干渉補正電極92を形成するのに使用する不活性層インクを実施例IIに記載した方法を用いて配合した。しかしながら、不活性層は、グルコースオキシダーゼのような酵素を触媒的に活性を有する量だけ含まない。5.0 グラムの結合剤樹脂を、2.0 グラムのBSA−PAC(実施例IIにおいて調製した)、0.25グラムのトリトン(登録商標)X−100界面活性剤および1.0 mlのアルベッソT(8101RS)シンナーとともに練磨を行なうことにより不活性層インクを調製した。この混合物にさらに0.4 グラムのペンテックス(登録商標)BSAを加えて練磨した。次いで、不活性層のペーストを導電金属片70の電極部分上にスクリーン印刷して干渉補正電極92を形成した。
ここで再度図1から12を参照する。実質的に本発明の一実施態様による構成部材および設計を含むセンサパッケージの適切な構成の1つを、以下の要素の組合せにより示す。
実質的に実施例I−IVにしたがって構成したプレーナグルコースセンサを全血により評価し、mg/dlで表したグルコースの濃度とナノアンペア(nA)で表したセンサの測定電流との関係を図17のグラフにプロットした。図17のグラフに示したように、このセンサの重要な特徴の1つは、グルコースの濃度がセンサにより測定された電流と線形の関係を示すことである。グルコースおよび酸素の両方が透過するセンサ膜94により、グルコースに対して化学量論的に過剰な酸素が、全血から膜に浸透し、その結果として、グラフの左端から右端まで線形の関係が得られると考えられている。
た。mモル/lで表したラクテートの濃度とナノアンペア(nA)で表したセンサにより測定された電流との関係を図18のグラフに示した。図18のグラフに示したように、このセンサの重要な特徴の1つは、ラクテートの濃度がセンサにより測定された電流と線形の関係を示すことである。ここで再度、ラクテートおよび酸素の両方が透過するセンサ膜により、ラクテートに対して化学量論的に過剰な酸素が、全血から膜に浸透して、その結果として、グラフの左端の約1.00mモル/lから右端の約20.0mモル/lまで線形の関係が得られると考えられている。
干渉補正電極92の効果を求めるために、この干渉補正電極を使用する場合としない場合の、グルコースの濃度に対するグルコースセンサの応答を記録して、図19のグラフに示した。電極92を設けて、中性物質であるアセトアミノフェンのような、センサの半透膜94を通って拡散し得る干渉物質のために調節した。
本発明の電気化学センサの寿命を測定するために、実質的に実施例I−IVにしたがって構成したグルコースセンサを、長期間に亘り多数の試料に対して用いた。このセンサを69日間に亘り試験した。この期間中に、2,250 の試料を評価した。グルコースの濃度が180 mg/dlである水溶液に対してnAで示した電流を
69日間に亘り測定した。図20に示すように、本発明のグルコースセンサは、69日間に亘り、および/または少なくとも2,250 の試料について、10nAより大きい応答を示した。
貯蔵期間を終了した後の最初の性能について、界面活性剤による後処理の効果を測定するために、センサを試験して、約83.0mg/dlから約470.0 mg/dlまでの範囲のグルコース濃度とナノアンペア(nA)で示したセンサにより測定した電流との間の関係を図23および24に示した。第1のセンサをトリトン(登録商標)X−100(前出)により後処理し、一方第2のセンサは後処理しなかった。ここでの評価を行なう前に、センサを室温で1週間に亘り貯蔵した。処理しなかったセンサにおいては、グルコースの濃度に対する応答が小さくて非線形であった(図24に示した拡大グラフからより明確に分かるように、処理しなかったセンサの応答は、約200.0 mg/dlのグルコース濃度を越えるとドリフトしてしまう)。これは、貯蔵中に膜が乾ききってしまうことによりウェットアップ(wetup )が遅くなった結果である。一方、処理したセンサにおいては、たった1時間のウェットアップ後に線形で完全なウェットアップ応答が達成された。
センサの応答の線形性に対する膜厚の影響を測定するために、2層の薄い膜 (約10ミクロン)、2層の厚い膜(約22.0ミクロン)、および4層の膜(約22.0ミクロンを別々に評価した。センサを試験して、mg/dlで示した水溶液中のグルコースの濃度とナノアンペア(nA)で示したセンサにより測定した電流との間の関係を図25にプロットした。ダウコーニング(ミシガン州、ミッドランド)からFC−61コーティングとして市販されているコロイド状シリカを約14.0重量パーセント含有する陰イオン安定化した水性の、末端がヒドロキシル基で終わったポリジメチルシロキサンエラストマーから多層膜の全てを調製した。図25から分かるように、多層の膜によりセンサの性能は改良され、このことは、グラフに示された線形応答により立証される。4層の膜と同じ厚さ(約22.0ミクロン)の厚い2層の膜は、出力が大きいが、グルコースの濃度に対する応答は線形ではなかった。4層の膜では、膜の欠点がなくなったために性能が改良されている。2層の膜は、酸素に対して過剰のグルコースが膜を通過してしまい、出力が大きいこと、並びに応答が線形ではないことの原因となる欠点がある。
白金化活性炭(PAC)材料に界面活性剤を含ませることの貯蔵に対する性能への効果を求めるために、室温で1日貯蔵した後と21日貯蔵した後のPAC中に界面活性剤を含ませない場合について、グルコースセンサの応答を評価した。センサを試験して、約500.0 mg/dlまでの範囲のグルコースの濃度と、ナノアンペア(nA)で示したセンサにより測定した電流との間の関係を図29にプロットした。図29から分かるように、トリトン(登録商標)X−100のような界面活性剤をPAC材料に加えなかった場合には、時間が経過するにつれてセンサの出力が低下する。図21は、最適化した多層膜についてであるが、図29と同様な効果を示している。
界面活性剤を膜94に加えることによるセンサの性能に対する影響を調べるために、グルコースセンサを試験し、約500.0 mg/dlまでの範囲のグルコースの濃度と、ナノアンペア(nA)で示したセンサにより測定した電流との関係を図31にプロットした。市販されているFC−61コーティング材料(前出)から実質的になるセンサ膜94をそれぞれのセンサに施した。一方のセンサには、ステパン社(イリノイ州、ノースフィールド)から得られる界面活性剤である、マコン(登録商標)10界面活性剤を含ませた。両方の膜とも、厚さが約11.0ミクロン厚である、2層のスピンキャスト膜である。膜に界面活性剤を添加することにより、グルコースの濃度に対する応答を大きくし、ウェットアップを改良することができる。
前記非導電性基体の第1の部分に接着した第1の導電性材料からなり、該第1の導電性材料の一部が絶縁コーティングにより覆われ、前記第1の導電性材料の一部が白金化炭素粉末粒子により担持された触媒的に活性を有する量の酵素を含む活性層により覆われ、前記白金化炭素粉末粒子が前記活性層全体に分布している作用電極、
前記非導電性基体の第2の部分に接着した第2の導電性材料からなり、該第2の導電性材料の一部が絶縁コーティングにより覆われ、前記第2の導電性材料の少なくとも一部が前記絶縁コーティングにより覆われずにいる対電極、
前記非導電性基体の第3の部分に接着した第3の導電性材料からなり、該第3の導電性材料の一部が絶縁コーティングにより覆われ、前記第3の導電性材料の少なくとも一部が前記絶縁コーティングにより覆われずにいる参照電極、および
前記作用電極を覆う半透膜
からなる多目的に使用できる固体電気化学センサ。
前記非導電性基体の第1の部分に接着した第1の導電性材料からなり、該第1の導電性材料の一部が絶縁コーティングにより覆われ、前記第1の導電性材料の一部が白金化炭素粉末粒子に固定化された触媒的に活性を有する量のグルコースオキシダーゼを含む活性層により覆われ、前記白金化炭素粉末粒子が前記活性層全体に実質的に均一に分布している作用電極、
前記非導電性基体の第2の部分に接着した第2の導電性材料からなり、該第2の導電性材料の一部が絶縁コーティングにより覆われ、前記第2の導電性材料の少なくとも一部が前記絶縁コーティングにより覆われずにいる対電極、
前記非導電性基体の第3の部分に接着した第3の導電性材料からなり、該第3の導電性材料の一部が絶縁コーティングにより覆われ、前記第3の導電性材料の少なくとも一部が前記絶縁コーティングにより覆われずにいる参照電極、および
前記電極を覆うグルコースおよび酸素透過性膜
からなる多目的に使用できる固体グルコースセンサ。
前記窪みの下にある固体平板電気化学センサ、および
前記窪みの境界面および前記固体平面電気化学センサに接触してこれらの間にシールを形成するガスケット
からなり、
前記ハウジング、前記ガスケット、および前記センサが試料室を形成することを特徴とするセンサパッケージ。
非導電性基体を選択し、
該非導電性基体上の第1の部分に導電性材料を付着させ、
該導電性材料上の一部に活性層を付着させて作用電極を形成し、ここで該活性層が白金化炭素粉末粒子に固定化された触媒的に活性を有する量の酵素を含み、前記白金化炭素粉末粒子が前記活性層全体に実質的に均一に分布しており、
前記非導電性基体上の第2の部分に第2の導電性材料を付着させて対電極を形成し、
前記非導電性基体上の第3の部分に第3の導電性材料を付着させて参照電極を形成し、 前記電極上に半透膜を付着させる
各工程からなることを特徴とする方法。
該第4の導電性材料上の一部に絶縁材料を付着させ、
前記第4の導電性材料上の他の一部分に不活性層を付着させて干渉補正電極を形成させる
各工程を含み、ここで前記不活性層が白金炭素粉末粒子に固定化した不活性タンパク質を含み、前記白金炭素粉末粒子が前記不活性層全体に実質的に均一に分布していることを特徴とする参考例31記載の方法。
前記活性層が、グルコースまたはラクテートの1つに対して反応性がある酵素と白金化炭素粉末粒子とからなり、それによって、グルコースまたはラクテートの1つが前記活性層にさらされるときに、前記活性層が前記グルコースまたはラクテート1つの量に比例した量のH2O2を形成させることができ、
前記外面保護層が前記酸素およびグルコースまたはラクテートの1つを移送させられるように含ませた添加剤を有するシリコーン化合物からなり、それによって、前記電極を用いて、グルコースまたはラクテートの濃度の1つを迅速かつ正確に測定できることを特徴とする電極。
速度調節部材が前記入口と出口との間の流路内に設けられた構造バリアからなり、この速度調節部材により前記試料室の前記流路に当る部分の断面積を減少させて、試験すべき流体が前記試料室を通過するときにこの流体の速度を実質的に安定化することを特徴とするセンサ。
前記リード線は該リード線に取り付けられた安定化棒により互いに離間されており、それによって前記リード線が、前記電気的接点に接触する適切な位置に確実に位置されることを特徴とするセンサ。
該非導電性基体の一部に接着した導電性材料を含む干渉電極であって、前記導電性材料の一部が絶縁コーティングにより覆われ、前記導電性材料の他の一部が白金化炭素粉末粒子に固定化された不活性タンパク質からなる不活性層により覆われ、前記白金化炭素粉末粒子が前記不活性層全体に実質的に均一に分布している干渉電極、そして
前記干渉電極を覆う半透膜
からなる電気化学センサ。
セルロースアセテートを蒸気圧の小さい溶媒と混合して粘度の大きいペーストを形成する工程を含むことを特徴とする方法。
(b) 蒸気圧の小さい溶媒
からなるセルロースアセテートペースト。
12 ハンドル部
14 本体
16 接触部
18 流体通路
20 ハウジング
22 窪み
24 境界面
26 ガスケット
28 センサ
30 センサパッド
32 接触リード線フレーム
34 リード線
36 ベース
42 センサ窪み
44 安定化棒
48 Oリング
50 スプリングチップ
54 試料室
56 入口
58 出口
60 速度調節部材
62 基体
64、66、68、70 導電性金属片
72 絶縁層
78、80、82、84 接触パッド
86、88、90、92 電極
94 活性層または保護膜
100 セルロースアセテート層
102 銀/塩化銀層
122 、124 側壁
126 前面穴
132 突起物
134 楕円形の穴
136 、138 突出面
Claims (2)
- (a) 白金化炭素粉末粒子を中性緩衝液と混合し、
(b) 上記工程(a) で得た緩衝化した白金化炭素粉末粒子をタンパク質と混合してタンパク質−白金化活性炭組成物を形成し、
(c) 上記工程(b) で得た前記タンパク質−白金化活性炭組成物を結合剤樹脂と混合し、
(d) 上記工程(c) で得た混合物を混練し、
(e) 上記工程(d) で得た混練生成物に酵素または不活性タンパク質のいずれかを加えて混合してペーストを形成する
各工程からなる電気化学電極用のペーストを形成する方法。 - (a) 白金化炭素粒子、
(b) タンパク質、
(c) 樹脂結合剤、および
(d) 酵素または不活性タンパク質のいずれか一方
からなる電気化学電極用のペースト。
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US08/266,824 US5494562A (en) | 1994-06-27 | 1994-06-27 | Electrochemical sensors |
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JP7161165A Division JPH08193969A (ja) | 1994-06-27 | 1995-06-27 | 電気化学センサ |
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Family
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JP7161165A Pending JPH08193969A (ja) | 1994-06-27 | 1995-06-27 | 電気化学センサ |
JP2004117096A Pending JP2004340949A (ja) | 1994-06-27 | 2004-04-12 | 電気化学センサ |
JP2004117093A Pending JP2004340948A (ja) | 1994-06-27 | 2004-04-12 | 電気化学センサ |
JP2004117091A Pending JP2004239915A (ja) | 1994-06-27 | 2004-04-12 | 電気化学センサ |
JP2004117095A Pending JP2004251913A (ja) | 1994-06-27 | 2004-04-12 | 電気化学センサ |
JP2004117094A Expired - Lifetime JP3869429B2 (ja) | 1994-06-27 | 2004-04-12 | 電気化学センサ |
JP2004117092A Pending JP2004340947A (ja) | 1994-06-27 | 2004-04-12 | 電気化学センサ |
JP2004117090A Pending JP2004245848A (ja) | 1994-06-27 | 2004-04-12 | 電気化学センサ |
Family Applications Before (4)
Application Number | Title | Priority Date | Filing Date |
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JP7161165A Pending JPH08193969A (ja) | 1994-06-27 | 1995-06-27 | 電気化学センサ |
JP2004117096A Pending JP2004340949A (ja) | 1994-06-27 | 2004-04-12 | 電気化学センサ |
JP2004117093A Pending JP2004340948A (ja) | 1994-06-27 | 2004-04-12 | 電気化学センサ |
JP2004117091A Pending JP2004239915A (ja) | 1994-06-27 | 2004-04-12 | 電気化学センサ |
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JP2004117092A Pending JP2004340947A (ja) | 1994-06-27 | 2004-04-12 | 電気化学センサ |
JP2004117090A Pending JP2004245848A (ja) | 1994-06-27 | 2004-04-12 | 電気化学センサ |
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EP (1) | EP0690134B1 (ja) |
JP (8) | JPH08193969A (ja) |
KR (1) | KR960001748A (ja) |
AT (1) | ATE188741T1 (ja) |
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CA (1) | CA2138856A1 (ja) |
DE (1) | DE69514427T2 (ja) |
DK (1) | DK0690134T3 (ja) |
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KR960001748A (ko) | 1996-01-25 |
ES2141893T3 (es) | 2000-04-01 |
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JP2004340947A (ja) | 2004-12-02 |
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AU704637B2 (en) | 1999-04-29 |
ATE188741T1 (de) | 2000-01-15 |
JP2004239915A (ja) | 2004-08-26 |
JP2004245848A (ja) | 2004-09-02 |
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