US5158868A — Method of sample analysis

Claim 1

Independent Claim 157 words

A method for measuring a constituent of interest of a biological fluid or the like comprising the steps of providing a reaction chamber that contains an immobilized enzyme capable of modifying a constituent of interest, providing a measuring system, placing a first portion of a biological fluid to be analyzed in said reaction chamber and concurrently exposing a second unmodified portion of said biological fluid to said measuring system to provide a first data output, oscillating said first biological fluid portion with bidirectional flow in said reaction chamber to facilitate modification by said immobilized enzyme of said constituent of interest in said biological fluid, then exposing said first portion of said biological fluid to be analyzed to said measuring system to provide a second data output, and modifying said second data output as a function of said first data output to provide an indication of the actual amount of said constituent of interest in said biological fluid.

Claim 2

Independent Claim 157 words

A method for measuring a constituent of interest of a biological fluid or the like comprising the steps of providing a reaction chamber that contains an immobilized enzyme capable of modifying a constituent of interest, providing a measuring system, placing a first portion of a biological fluid to be analyzed in said reaction chamber and concurrently exposing a second unmodified portion of said biological fluid to said measuring system to provide a first data output, oscillating said first biological fluid portion with bidirectional flow in said reaction chamber to facilitate modification by said immobilized enzyme of said constituent of interest in said biological fluid, then exposing said first portion of said biological fluid to be analyzed to said measuring system to provide a second data output, and modifying said second data output as a function of said first data output to provide an indication of the actual amount of said constituent of interest in said biological fluid.

Claim 3

Dependent Claim 76 words

The method of claim 1 wherein said measuring system includes an analysis region and said analysis region and said reaction chamber are connected together in a series flow path, and said first and second portions of said biological fluid are placed in said reaction chamber and exposed to said measuring system by flowing said biological fluid to be analyzed through said reaction chamber at a flow rate of at least about one hundred microliters per second.

Claim 4

Dependent Claim 24 words

The method of claim 1 wherein said constituents of interest are urea and glucose, and two different enzymes are coimmobilized in said reaction chamber.

Claim 5

Dependent Claim 31 words

The method of claim 3 wherein said two different coimmobilized enzymes are glucose oxidase and urease and said measuring system includes an ammonium ion selective electrode and an oxygen sensing electrode.

Claim 6

Independent Claim 198 words

A method of measuring a substance capable of being enzymatically converted to a detectable ion with compensation for potassium ion interference comprising the steps of placing a sample containing a substance capable of being enzymatically converted to a detectable ion in a chamber comprising an immobilized enzyme capable of converting said substance to said detectable ion and detecting means for detecting said detectable ion; contacting said sample with said detecting means before said immobilized enzyme has converted said substance to said detectable ion and measuring the output of said detecting means to provide a potassium ion interference (pre-enzymatic reaction) output; oscillating said sample in said chamber in a bidirectional flow mode interval to facilitate conversion by said immobilized enzyme of said substance in said sample to said detectable ion; contacting said sample with said detecting means after said immobilized enzyme has converted said substance to said detectable ion and measuring the output of said detecting means to provide an apparent substance amount output; and modifying said post-enzymatic reaction output as a function of said pre-enzymatic reaction output to provide an indication of the actual amount of a substance in said sample using a form of the Nicolsky equation.

Claim 7

Independent Claim 198 words

A method of measuring a substance capable of being enzymatically converted to a detectable ion with compensation for potassium ion interference comprising the steps of placing a sample containing a substance capable of being enzymatically converted to a detectable ion in a chamber comprising an immobilized enzyme capable of converting said substance to said detectable ion and detecting means for detecting said detectable ion; contacting said sample with said detecting means before said immobilized enzyme has converted said substance to said detectable ion and measuring the output of said detecting means to provide a potassium ion interference (pre-enzymatic reaction) output; oscillating said sample in said chamber in a bidirectional flow mode interval to facilitate conversion by said immobilized enzyme of said substance in said sample to said detectable ion; contacting said sample with said detecting means after said immobilized enzyme has converted said substance to said detectable ion and measuring the output of said detecting means to provide an apparent substance amount output; and modifying said post-enzymatic reaction output as a function of said pre-enzymatic reaction output to provide an indication of the actual amount of a substance in said sample using a form of the Nicolsky equation.

Claim 8

Dependent Claim 17 words

The method of claim 5 wherein said substance comprises urea and said detectable ion comprises ammonium ion.

Claim 9

Dependent Claim 23 words

The method of claim 5 wherein said detecting means comprises an ion selective electrode and a reference electrode, said ion selective and reference

Claim 10

Independent Claim 240 words

A method of measuring a substance capable of being enzymatically converted to a detectable ion with compensation for potassium ion interference comprising the steps of placing a sample containing a substance capable of being enzymatically converted to a detectable ion in a chamber comprising an immobilized enzyme capable of converting said substance to said detectable ion and detecting means for detecting said detectable ion; contacting said sample with said detecting means before said immobilized enzyme has converted said substance to said detectable ion and measuring the output of said detecting means to provide a potassium ion interference (pre-enzymatic reaction) output; oscillating said sample in said chamber with bidirectional flow to facilitate conversion by said immobilized enzyme of said constituent of interest in said sample to said detectable ion; contacting said sample with said detecting means after said enzyme has converted said substance to said detectable ion and measuring the output of said detecting means to provide an apparent substance amount (post-enzymatic reaction) output; and modifying said post-enzymatic reaction output as a function of said pre-enzymatic reaction output to provide an indication of the actual amount of a substance in said sample. electrodes are spaced from one another by a flow path portion located so that said reference electrode is downstream of and spaced from said ion selective electrode by a flow path volume greater than the displacement volume that sample material is moved during said bidirectional sample flow mode interval.

Claim 11

Independent Claim 240 words

A method of measuring a substance capable of being enzymatically converted to a detectable ion with compensation for potassium ion interference comprising the steps of placing a sample containing a substance capable of being enzymatically converted to a detectable ion in a chamber comprising an immobilized enzyme capable of converting said substance to said detectable ion and detecting means for detecting said detectable ion; contacting said sample with said detecting means before said immobilized enzyme has converted said substance to said detectable ion and measuring the output of said detecting means to provide a potassium ion interference (pre-enzymatic reaction) output; oscillating said sample in said chamber with bidirectional flow to facilitate conversion by said immobilized enzyme of said constituent of interest in said sample to said detectable ion; contacting said sample with said detecting means after said enzyme has converted said substance to said detectable ion and measuring the output of said detecting means to provide an apparent substance amount (post-enzymatic reaction) output; and modifying said post-enzymatic reaction output as a function of said pre-enzymatic reaction output to provide an indication of the actual amount of a substance in said sample. electrodes are spaced from one another by a flow path portion located so that said reference electrode is downstream of and spaced from said ion selective electrode by a flow path volume greater than the displacement volume that sample material is moved during said bidirectional sample flow mode interval.

Claim 12

Dependent Claim 64 words

The method of claim 8 further comprising the step of calibrating said detecting means using first and second calibrators having known concentrations of potassium ions and said substance by contacting said first and second calibrators individually with said detecting means and measuring the output of said detecting means before and after said enzyme has converted said substance in said calibrators to said detectable ion.

Claim 13

Dependent Claim 119 words

The method of claim 9 wherein said substance is urea and the actual concentration of urea in said sample is determined by effectively subtracting said pre-enzymatic reaction data from said post enzymatic reaction data using the following form of equation: conc..sub.UN =antilog [(E.sub.S -E.sub.cal 1)(1/S)] * (Cal 1 UN +kCal 1k )-(k conc. K) where conc. UN =actual concentration of urea in the sample; E s =post-enzymatic reaction measurement for the sample in millivolts; E cal1 =post-enzymatic reaction measurement for calibrator 1 in millivolts; Cal 1 UN =actual concentration of urea in calibrator 1 (known); Cal 1k =actual concentration of potassium in calibrator 1 (known); k=potassium selectivity factor for the electrode; and conc. K=concentration of potassium in the sample.

Claim 14

Dependent Claim 20 words

The method of claim 10 wherein said ion selective electrode includes nonactin ionophore, and said immobilized enzyme includes urease enzyme.

Claim 15

Dependent Claim 28 words

The method of claim 8 that further includes the concurrent measurement of a second substance in said sample, and a second enzyme is coimmobilized in said reaction chamber.

Claim 16

Dependent Claim 48 words

The method of claim 12 wherein said detecting means and said chamber are connected together in a series flow path, and said sample to be analyzed is flowed through said chamber to said detecting means at a flow rate of at least about one hundred microliters per second.

Claim 17

Dependent Claim 22 words

The method of claim 13 wherein said second substance in said sample is glucose and said coimmobilized second enzyme is glucose oxidase.

Claim 18

Dependent Claim 20 words

The method of claim 14 wherein said ion selective electrode includes nonactin ionophore, and the other coimmobilized enzyme is urease.

Annuity Stage	Window Opens	Payment Deadline	Surcharge Period Until	Estimated Fee	Status
3.5 Years (Year 4)	1995-10-27	1996-04-27	1996-10-27	$2000.00	Window Closed / Lapsed
7.5 Years (Year 8)	1999-10-27	2000-04-27	2000-10-27	$3760.00	Window Closed / Lapsed
11.5 Years (Year 12)	2003-10-27	2004-04-27	2004-10-27	$7700.00	Window Closed / Lapsed

Method of sample analysis

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