Rate sensing batch analysis method and enzyme used therein

• US 4,045,296 A
• Filed: 12/29/1975
• Issued: 08/30/1977
• Est. Priority Date: 08/09/1972
• Status: Expired due to Term

• Current Assignee: Beckman Instruments, Inc. (Danaher Corporation)
• Original Assignee: Beckman Instruments, Inc. (Danaher Corporation)
• Inventors: Sternberg, James C.
• Primary Examiner(s): Tanenholtz, Alvin E.
• Assistant Examiner(s): Fan, C. A.
• Application Number:
  US05/645,009
• Time in Patent Office: 610 Days
• Field of Search: 195/103.5 R, 195/103.5 C, 204/195 B, 204/1 E
• US Class Current: 435/11
• CPC Class Codes: G01N 27/3271 Amperometric enzyme electrodes for analytes in body fluids, e.g. glucose in blood amperometry per se G01N27/49; aspects concerning the enzyme reagent C12Q1/001
• CPC Class Codes: Y10S 435/807 Gas detection apparatus

Abstract

A portable, automated chemical analyzer having a polarographic oxygen electrode submerged in a batch of stirred solution. The signal from the electrode is directly differentiated and amplified to produce a signal proportional to the time rate of change of oxygen concentration. The membrane of the polarographic sensor is stretched very tightly over the cathode surface to provide a high signal-to-noise ratio. Methods for analyzing cholesterol-cholesterol oxidase and various other enzyme systems by the polarographic electrode oxygen sensing apparatus include the steps of converting the sensed signal into time rate of change of oxygen concentration and recording the maximum rate of change of oxygen concentration.

Claims

1. A method of determining the cholesterol concentration in a cholesterol containing sample comprising the steps of:

oxidizing the cholesterol present in the sample in an oxygen saturated aqueous solution by means of cholesterol oxidase in the presence of a sensor which serves to monitor a property or characteristic of oxygen in said solution related to the oxygen concentration thereof;

generating a first electrical signal related to said oxygen concentration;

differentiating said first electrical signal to produce an output signal proportional to the instantaneous time rate of change of oxygen concentration; and

measuring said output signal to determine said cholesterol concentration.