Claims Analysis
Total Claims: 8
Claim 1
Independent Claim
177 words
A TOTENTIOMETRIC METHOD FOR SELECTIVELY DETERMING THE CONCENTRATION OF ALKALINE PHOSPHATASE OF AN AQUEOUS FLUID SAMPLE, COMPRISING, A. DILUTING THE AQUEOUS FLUID SAMPLE WITH A BUFFERED AQUEOUS SOLUTION HAVING A PH IN THE RANGE OF ABOUT 8.5 TO ABOUT 9, B. PROVIDING A POTENTIOMETER ELECTRICALLY CONNECTED IN SERIES TO A D-SERINE SPECIFIC ELECTRODE AND A REFERENCE ELECTRODE, WHEREIN THE D-SERINE SPECIFIC ELECTRODE IS PROVIDED WITH AN AMMONIUM ION-SENSITIVE MEMBRANE IN CONTACT WITH AND SUBSTANTIALLY ENCAPSULATED BY A COATING OF INSOLUBILIZED D-SERINE DEAMINASE, SAID COATING CONTANING A CATALYTICALLY EFFECTIVE AMOUNT OF D-SERINE DEAMINASE AND BEING PERMEABLE TO BOTH THE AQUEOUS FLUID SAMPLE AND PRODUCTS OF THE REACTION OF THE AQUEOUS FLUID SAMPLE AND THE D-SERINE DEAMINASE, C. CONTACTING THE D-SERINE SPECIFIC ELECTRODE AND THE REFERENCE ELECTRODE WITH DILUTED SAMPLE, D. RECORDING THE IONIC BACKGROUND ACTIVITY OF THE SAMPLE E. ADDING TO THE DILUTED SAMPLE ABOUT 5X10**4 TO ABOUT 5X10**1 MOLES D-PHOSPHOSERINE PER LITER OF DILUTED SAMPLE, F. AND RECORDING THE INCREMENTAL CHANGE IN POTENTIAL AT REGULAR INTERVALS FOR A PERIOD RANGING FROM ABOUT 5 TO ABOUT 15 MINUTES.
Claim 2
Dependent Claim
177 words
A TOTENTIOMETRIC METHOD FOR SELECTIVELY DETERMING THE CONCENTRATION OF ALKALINE PHOSPHATASE OF AN AQUEOUS FLUID SAMPLE, COMPRISING, A. DILUTING THE AQUEOUS FLUID SAMPLE WITH A BUFFERED AQUEOUS SOLUTION HAVING A PH IN THE RANGE OF ABOUT 8.5 TO ABOUT 9, B. PROVIDING A POTENTIOMETER ELECTRICALLY CONNECTED IN SERIES TO A D-SERINE SPECIFIC ELECTRODE AND A REFERENCE ELECTRODE, WHEREIN THE D-SERINE SPECIFIC ELECTRODE IS PROVIDED WITH AN AMMONIUM ION-SENSITIVE MEMBRANE IN CONTACT WITH AND SUBSTANTIALLY ENCAPSULATED BY A COATING OF INSOLUBILIZED D-SERINE DEAMINASE, SAID COATING CONTANING A CATALYTICALLY EFFECTIVE AMOUNT OF D-SERINE DEAMINASE AND BEING PERMEABLE TO BOTH THE AQUEOUS FLUID SAMPLE AND PRODUCTS OF THE REACTION OF THE AQUEOUS FLUID SAMPLE AND THE D-SERINE DEAMINASE, C. CONTACTING THE D-SERINE SPECIFIC ELECTRODE AND THE REFERENCE ELECTRODE WITH DILUTED SAMPLE, D. RECORDING THE IONIC BACKGROUND ACTIVITY OF THE SAMPLE E. ADDING TO THE DILUTED SAMPLE ABOUT 5X10**4 TO ABOUT 5X10**1 MOLES D-PHOSPHOSERINE PER LITER OF DILUTED SAMPLE, F. AND RECORDING THE INCREMENTAL CHANGE IN POTENTIAL AT REGULAR INTERVALS FOR A PERIOD RANGING FROM ABOUT 5 TO ABOUT 15 MINUTES.
Claim 3
Dependent Claim
21 words
The potentiometric method as defined in claim 1, wherein the D-serine specific electrode is referenced to a constant potential reference electrode.
Claim 4
Dependent Claim
22 words
The potentiometric method as defined in claim 2, wherein the diluted aqueous fluid sample is pre-treated with a strongly acidic cation-exchange resin.
Claim 5
Dependent Claim
27 words
The potentiometric method as defined in claim 1, wherein the diluted sample is periodically agitated after addition of D-phosphoserine during recordation of changes in enzyme electrode potential.
Claim 6
Dependent Claim
175 words
A potentiometric method for selectively determining the concentration of alkaline phosphatase in an aqueous fluid sample having a pH in the range of about 8.5 to 9, the method comprising the steps of: a. providing a potentiometer electrically connected to an enzyme electrode and a variable potential reference electrode, in which the enzyme electrode is provided with an ammonium ion-sensitive membrane in contact with and substantially encapsulated by a coating of immobilized enzyme, said coating containing a catalytically effective amount of D-serine deaminase, said coating being permeable to both the aqueous fluid sample and products of the reaction of the aqueous fluid sample and the D-serine deaminase; and in which the variable potential reference electrode is provided with an uncoated ammonium ion-sensitive membrane of substantially the same ionic sensitivity and selectivity as the enzyme electrode; and b. contacting the ammonium ion-sensitive membranes of both the enzyme electrode and the variable potential electrode with the sample of aqueous fluid and about 5 X 10 4 to 5 X 10 1 moles D-phosphospherine per liter of sample.
Claim 7
Dependent Claim
175 words
A potentiometric method for selectively determining the concentration of alkaline phosphatase in an aqueous fluid sample having a pH in the range of about 8.5 to 9, the method comprising the steps of: a. providing a potentiometer electrically connected to an enzyme electrode and a variable potential reference electrode, in which the enzyme electrode is provided with an ammonium ion-sensitive membrane in contact with and substantially encapsulated by a coating of immobilized enzyme, said coating containing a catalytically effective amount of D-serine deaminase, said coating being permeable to both the aqueous fluid sample and products of the reaction of the aqueous fluid sample and the D-serine deaminase; and in which the variable potential reference electrode is provided with an uncoated ammonium ion-sensitive membrane of substantially the same ionic sensitivity and selectivity as the enzyme electrode; and b. contacting the ammonium ion-sensitive membranes of both the enzyme electrode and the variable potential electrode with the sample of aqueous fluid and about 5 X 10 4 to 5 X 10 1 moles D-phosphospherine per liter of sample.
Claim 8
Dependent Claim
23 words
A method as defined in claim 5 in which the further step is provided of measuring the generated electrical potential on the potentiometer.