Apparatus for measuring substrate concentrations

• US 4,005,002 A
• Filed: 08/01/1974
• Issued: 01/25/1977
• Est. Priority Date: 08/06/1973
• Status: Expired due to Term

• Current Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Original Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Inventors: Higelin, Jean-Claude, Racine, Philippe, Engelhardt, Roland, Mindt, Wolfgang
• Primary Examiner(s): Tung, T.
• Application Number:
  US05/493,583
• Time in Patent Office: 908 Days
• Field of Search: 204/195 B, 204/195 R, 204/195 P, 23/230 A, 23/253 A, 195/103.5 R, 195/103.5 C
• US Class Current: 204/403.1
• CPC Class Codes: C12Q 1/004 mediator-assisted
• CPC Class Codes: Y10S 435/817 Enzyme or microbe electrode

Abstract

The concentration of substrates of enzyme reactions is measured by determining the current produced at a platinum or gold electrode by oxidizing an acceptor utilizing a measuring device comprising a measuring cell, an enzyme electrode in contact with the fluid sample and containing a platinum or gold electrode, an enzyme layer and a reference electrode in contact with the fluid sample. In this manner important physiological parameters, such as the concentration of lactate and glucose in biological fluids, can be measured quickly and accurately.

Claims

1. A device for measuring the concentration of substrates of enzyme reactions occurring during intermediate metabolism, more particularly the concentration of lactate and glucose in fluid biological samples, including a measuring cell formed as a tubular chamber for receiving the fluid sample, an enzyme electrode containing a platinum or gold electrode, an enzyme layer and a semi-permeable membrane covering the enzyme layer;

a reference electrode;

said tubular chamber being placed between the contact surfaces of said electrodes; and

an electrical circuit for measuring the current which flows through the platinum or gold electrode,

which device is characterized in that it comprises means for locally regulating the temperature of the enzyme layer, a hydraulic device for alternately filling the measuring cell with an acceptor-containing buffer solution or a fluid sample, and said electrical current measuring circuit includes an electronic circuit which, after the measuring cell has been filled with a fluid sample, determines the peak value of the current flowing through the platinum or gold electrode, the peak value, being representative of the concentration of substrate to be measured.

2. The device of claim 1 characterized in that the enzyme electrode comprises a temperature sensor which is thermally coupled to the platinum or gold electrode and which provides a temperature-regulating unit with a signal representing the actual temperature of the platinum or gold electrode and a Peltier cell which is thermally coupled to the platinum or gold electrode and is controlled by the temperature-regulating unit so as to regulate the temperature of the platinum or gold electrode and of the enzyme-containing layer in contact therewith.

3. The device of claim 1 wherein the reference electrode contains components which prevent ions liberated at the reference electrode from reaching the enzyme electrode.

4. The device of claim 1 wherein said hydraulic device comprises a first container holding a buffer solution containing an acceptor, a distributor which is connected to the first container and to the measuring cell system and is used to transfer buffer solution from the first container into the measuring cell or to introduce a fluid sample into the measuring cell;

a second container for receiving waste solutions sucked from the measuring cell;

a vacuum pump connected to the second container for producing a negative pressure therein so that the buffer solution can flow from the first container into the measuring cell or from the measuring cell into the second container; and

an air valve connected to the second container so as to overcome the negative pressure in the second container produced by the vacuum pump and thus interrupt the flow caused by the difference in pressure.

5. The device of claim 4 wherein there is an automatic control unit which cyclically controls the hydraulic device.

6. The device of claim 5 wherein there is an indicating means which is controlled by said automatic control unit and indicates that the device is ready to receive a fluid sample.

7. The device of claim 1 wherein said means for determining said peak value is an electric indicating device comprising a current amplifier connected to the reference electrode and the enzyme electrode;

a d.c. source connected between the enzyme electrode and the reference electrode;

an ammeter which is connected to one input of the current amplifier and gives a measurement corresponding to the current flowing through the platinum or gold electrode; and

a window discriminator which is connected in parallel to the ammeter and is used to determine the peak value of the current flowing through the platinum or gold electrode.

8. The device of claim 7 wherein there is an indicating device which is controlled by the window discriminator for indicating the point of time when the current flowing through the enzyme electrode reaches its peak value.

9. The device of claim 8 wherein an automatic control unit controls said electric indicator device.