Abstract A continuous method and apparatus for the determination of urea in a biological fluid such as blood, for example, is disclosed. In a continuously flowing system, the biological sample is introduced into a buffered carrier solution of low electrical conductance and is caused to flow into a chamber containing an immobilized enzyme which catalyzes a specific reaction which changes the electrical properties of the solution by increasing the dissociated ion concentration therein. This increase in dissociated ion concentration is directly proportional to the original concentration of urea in the biological sample; and any one of the several conventional electrical measurements may be made to determine the increase in such concentration or the rate of increase in such concentration and thereby determine the concentration of urea in the original biological sample. The buffered carrier solution containing the products of the enzyme-catalyzed reaction is reclaimed by circulating it through a deionization, purification stage in which the solution conductivity is substantially returned to the prereaction value. The carrier solution is recirculated and used for processing additional samples. The enzyme, urease, is immobilized in a manner which allows many tests to be run without any significant loss thereof. The savings in enzyme requirements and the savings accomplished by recirculating the buffered carrier solution greatly reduces the cost of analysis.
A continuous method for determining the concentration of a component of interest in a sample wherein the detection of the concentration of charge carriers of an ionic indicating product species of an enzyme-catalyzed reaction is indicative of the concentration of said component of interest, said method comprising the steps of: combining said sample with a substantially ionically inactive carrier solution containing an organic buffer; flowing the combined solution through a quantity of an enzyme selected to catalyze the desired reaction, wherein said enzyme is immobilized in a manner which allows free contact between the solution components and the enzyme molecules occasioning substantially no loss of enzyme thereby; measuring the increase in conductance of said combined solution; generating an output indicative of the concentration of said component of interest; reclaiming said carrier solution after said enzyme catalyzed reaction by reducing the conductance of the reacted solution substantially to that existing before said reaction; and recycling said reclaimed carrier solution to be reused with subsequent samples.
Claim 2
Dependent Claim
21 words
The method of claim 1 wherein said output is generated by measuring the electrical conductance of said solution after said reaction.
Claim 3
Dependent Claim
38 words
The method of claim 1 wherein said output is generated by measuring the conductance of said solution both before and after said enzyme-catalyzed reaction, the difference therebetween being indicative of the concentration of said ionic indicating product species.
Claim 4
Dependent Claim
26 words
The method of claim 1 wherein said output is generated by measuring the rate of change of the electrical conduction of said solution during said reaction.
Claim 5
Dependent Claim
20 words
The method of claim 1 wherein said reclaiming of said carrier solution is achieved by substantially de-ionizing the reacted solution.
Claim 6
Dependent Claim
18 words
The method of claim 1 wherein said component of interest is urea and said immobilized enzyme is urease.
Claim 7
Dependent Claim
164 words
A continuous flow-through analytical apparatus for determining the concentration of a component of interest in a sample wherein the detection of the concentration of charge carriers of an ionic indicating product species of an enzyme-catalyzed reaction is indicative of the concentration of the component of interest, said apparatus comprising: a first chamber including an amount of an enzyme immobilized and disposed therein in a manner which allows free contact between said enzyme and a solution flowing therethrough, means for introducing a substantially ionically inactive, carrier solution containing an organic buffer into said chamber; means for introducing a sample into said carrier solution; sensor means for detecting the increase in said charge carriers in said combined solution; means for generating an output from said sensor related to the concentration of said species of interest; means for reclaiming said carrier solution after said reaction by reducing the conductance of said carrier solution substantially to that existing before said reaction; and means for recirculating said reclaimed solution.
Claim 8
Dependent Claim
23 words
The apparatus of claim 7 wherein said sensor means comprises a pair of electrical conductance measuring electrodes placed downstream of said first chamber.
Claim 9
Dependent Claim
27 words
The apparatus of claim 8 further comprising a second pair of conductance-measuring electrodes disposed in said apparatus at a point before said solution enters said first chamber.
Claim 10
Dependent Claim
31 words
The apparatus of claim 7 wherein said sensor means comprises a pair of conductance-measuring electrodes placed in said first chamber to measure the rate of production of said ionic charge carriers.