Electrochemical detection of analytes
First Claim
1. A method of detecting an analyte wherein the analyte is either (a) a substrate which is oxidisable by means of an oxidase with the generation of hydrogen peroxide, the quantity of hydrogen peroxide being dependent on the quantity of analyte, or said analyte is convertible into a said oxidisable substrate, or (b) said oxidase;
- said method comprising;
(a) providing component A which comprises (a) said oxidase if the analyte is said oxidisable substrate, or (b) said oxidisable substrate if the analyte is said oxidase;
providing component B which comprises a peroxidase capable of oxidising a mediator comprising 2,2′
-azino-bis-(3-ethylbenzthiazoline-6-sulphonic acid) (“
ABTS”
) with concomitant reduction of hydrogen peroxide; and
providing an electrochemical cell comprising a redox electrode connected to a counter-electrode;
(b) allowing a sample suspected to contain said analyte to contact component A so that analyte present in the sample interacts with component A with the generation of hydrogen peroxide;
(c) allowing the generated hydrogen peroxide to contact the reduced form of said peroxidase in the presence of ABTS so that said hydrogen peroxide is reduced and said ABTS is oxidised by said peroxidase to form ABTSOX; and
(d) Allowing said ABTSOX to contact said redox electrode at which it undergoes reduction to ABTS, causing an electrical current to flow in said cell; and
(e) measuring said current to provide an output signal indicative of the presence of analyte.
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Abstract
An analyte is determined electrochemically indirectly. In a first step an oxidase and an oxidisable substrate, one of which is the analyte or derived therefrom, interact and generate hydrogen peroxide. In a second step a peroxidase (especially horse-radish peroxidase) reduces the hydrogen peroxide and concomitantly oxidises a mediator, preferably 2,2′-azino-bis(3-ethyl=benzthiazoline-6-sulfonic acid, ABTS) to an oxidised form (ABTSOX). The oxidised mediator is then reduced at an electrode and the consequent current is measured. A preferred sensor format uses a carbon electrode screen-printed onto a substrate and overlaid with one or more layers containing the enzymes and other components.
241 Citations
21 Claims
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1. A method of detecting an analyte wherein the analyte is either (a) a substrate which is oxidisable by means of an oxidase with the generation of hydrogen peroxide, the quantity of hydrogen peroxide being dependent on the quantity of analyte, or said analyte is convertible into a said oxidisable substrate, or (b) said oxidase;
- said method comprising;
(a) providing component A which comprises (a) said oxidase if the analyte is said oxidisable substrate, or (b) said oxidisable substrate if the analyte is said oxidase;
providing component B which comprises a peroxidase capable of oxidising a mediator comprising 2,2′
-azino-bis-(3-ethylbenzthiazoline-6-sulphonic acid) (“
ABTS”
) with concomitant reduction of hydrogen peroxide; and
providing an electrochemical cell comprising a redox electrode connected to a counter-electrode;
(b) allowing a sample suspected to contain said analyte to contact component A so that analyte present in the sample interacts with component A with the generation of hydrogen peroxide;
(c) allowing the generated hydrogen peroxide to contact the reduced form of said peroxidase in the presence of ABTS so that said hydrogen peroxide is reduced and said ABTS is oxidised by said peroxidase to form ABTSOX; and
(d) Allowing said ABTSOX to contact said redox electrode at which it undergoes reduction to ABTS, causing an electrical current to flow in said cell; and
(e) measuring said current to provide an output signal indicative of the presence of analyte. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20)
- said method comprising;
-
21. (Canceled).
Specification