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Enzyme Electrode

Enzyme electrodes work by immobilizing an enzyme like urease in a gel coating on an ion-selective electrode. Urease catalyzes the breakdown of urea into ammonium ions. A urea electrode dipped in a urea solution will allow urea to diffuse into the gel and be broken down by urease. The produced ammonium ions diffuse to the electrode surface and are detected, resulting in a potential reading proportional to the urea concentration. Enzyme electrodes can be used to measure analytes like urea, glucose, and ethanol through coupling selective enzyme reactions with amperometric electrode processes.

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2K views10 pages

Enzyme Electrode

Enzyme electrodes work by immobilizing an enzyme like urease in a gel coating on an ion-selective electrode. Urease catalyzes the breakdown of urea into ammonium ions. A urea electrode dipped in a urea solution will allow urea to diffuse into the gel and be broken down by urease. The produced ammonium ions diffuse to the electrode surface and are detected, resulting in a potential reading proportional to the urea concentration. Enzyme electrodes can be used to measure analytes like urea, glucose, and ethanol through coupling selective enzyme reactions with amperometric electrode processes.

Uploaded by

Atheer ab
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
  • Enzyme Electrodes
  • Classification of ISEs
  • Enzyme Electrodes Overview
  • Hydrolysis of Urea
  • Measurement of Urea Hydrolysis
  • Preparation of Enzyme Electrode
  • Enzyme-Based Sensors
  • Advantages and Disadvantages
  • Application of Enzyme Electrodes

Enzyme Electrodes

Classification of ISEs :

■ Glass membrane electrodes


■ Solid- state membrane electrodes
■ liquid membrane electrodes
■ plastic membrane electrodes
■ Special arragements such as }gas- sensitive
electrodes {, }Enzyme electrodes *Urea*{ .
Enzyme electrodes:

■ Enzyme are proteins that catalyze specific reactions to a high degree of


specificity .
■ Enzyme reacts with a specific substance, and the product of this reaction
(usually H+ or OH−) is detected by a true ion-selective electrode, such as a
pH-selective electrodes.
■ All these reactions occur inside a special membrane which covers the true
ion-selective electrode
■ Exmples Hydrolysis of the substrate urea in the presence of the enzyme urease :
■ Urease : An enzyme capable of cracking urea into ammonium .
■ When the electrode is dipped into a solution containing urea , the following
reaction occurs to yield N𝐻4+ .
𝑈𝑟𝑒𝑎𝑠𝑒
■ 𝑁𝐻2 𝐶𝑂𝑁𝐻2 +2𝐻2 O + 𝐻 +
2N𝐻4+ + HC𝑂3−
■ The N𝐻4+ is formed during the reaction is measured at the ammonium –
selective electrode .
■ the potential due to production of the N𝐻4+ ion following the equation :
E=𝐸 + 0.059 log (N𝐻4+ )
Δ𝐸
 A plot of versus concentration of enzyme or substrate always gave a straight
Δ𝑡
line in all the enzyme .
the hydrolysis of the substrate urea in the presence of the
enzyme urease:
𝑈𝑟𝑒𝑎𝑠𝑒
𝑁𝐻2 𝐶𝑂𝑁𝐻2 +2𝐻2 O + 𝐻 + 2N𝐻4+ + HC𝑂3−

To measure urea,NH2CONH2,in solution you can use an electrode


selective to N𝐻4+ that is coated with a gel containing the enzyme
urease.

The gel is held in place around the electrode by means of a nylon net.

Urease catalyzes the decomposition of urea to ammonium ion


Enzyme Electrode
• A urea electrode can be prepared by immobilizing
urease in a gel and coating it on the surface of a
cation-sensitive-type glass electrode(that responds to
monovalent cations).

• When the electrode is dipped in a solution containing


urea, the urea diffuses into the gel layer and the
enzyme catalyzes its hydrolysis to form ammonium
ion.
• The ammonium ions diffuse to the surface of the
electrode where they are sensed by the cation-
sensitive glass to give a potential reading.

• After about 30 to 60 seconds, a steady- state reading


is reached which, over a certain working range, is a
linear function of the logarithm of the urea
concentration.
Advantages and disadvantages :
■ Advantages :
1. The potential exists for developing electrodes
2. The potential sensed by the electrode in the solution after hydrolysis is determined
by the amount of ammonium ion produced and therefore is given by the
concentration of urea.
■ Disadvantages :
1. Gels must be replaced regularly
2. Each measurement results in cumulative response
3. The response is time dependent
Application:

■ coupling of appropriate amperometric electrode processes with enzyme system


eg ; urease result in a of simplification operation
 useful for monitoring clinical , environmental , food samples
 For determination of glucose in blood ( glucose sensors) .
 for amperometric sensing of ethanol (ethanol sensors ).
 For sensing urea in the presence of urease enzyme (urea electrode ) .

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