A kinetic method for determination of free vanadium(IV) and (V) at trace level concentrations

A kinetic method based on alkaline phosphatase has been developed to measure free trace levels of vanadium(IV) and (V). The method involves measuring the rate of the alkaline phosphatase-catalyzed hydrolysis of p-nitrophenyl phosphate with (Vi) and without (Vo) a competitive inhibitor in the assay....

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Published in:Analytical biochemistry Vol. 188; no. 1; p. 53
Main Authors: Crans, D C, Gottlieb, M S, Tawara, J, Bunch, R L, Theisen, L A
Format: Journal Article
Language:English
Published: United States 01-07-1990
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Abstract A kinetic method based on alkaline phosphatase has been developed to measure free trace levels of vanadium(IV) and (V). The method involves measuring the rate of the alkaline phosphatase-catalyzed hydrolysis of p-nitrophenyl phosphate with (Vi) and without (Vo) a competitive inhibitor in the assay. Michaelis-Menten kinetics for a competitive inhibitor was used to express the relationship between Vo/Vi and the inhibitor concentration. Measuring both Vo and Vi thus yields a Vo/Vi ratio that allows calculation of the competitive inhibitor concentration. Determination of free vanadium in complex fluids can be accomplished by comparing the ratio of rates of p-nitrophenyl phosphate hydrolysis with and without a sequestering agent to the ratios of rates measured on addition of a known vanadium concentration. Free vanadium(V) can conveniently be measured from 10(-7) to 10(-5) M and free vanadium(IV) can be measured at 10(-8) M and above. The error limits on the vanadium determinations range from +/- 3 to +/- 12% of the concentration under investigation depending on the conditions under which the assay was conducted.
AbstractList A kinetic method based on alkaline phosphatase has been developed to measure free trace levels of vanadium(IV) and (V). The method involves measuring the rate of the alkaline phosphatase-catalyzed hydrolysis of p-nitrophenyl phosphate with (Vi) and without (Vo) a competitive inhibitor in the assay. Michaelis-Menten kinetics for a competitive inhibitor was used to express the relationship between Vo/Vi and the inhibitor concentration. Measuring both Vo and Vi thus yields a Vo/Vi ratio that allows calculation of the competitive inhibitor concentration. Determination of free vanadium in complex fluids can be accomplished by comparing the ratio of rates of p-nitrophenyl phosphate hydrolysis with and without a sequestering agent to the ratios of rates measured on addition of a known vanadium concentration. Free vanadium(V) can conveniently be measured from 10(-7) to 10(-5) M and free vanadium(IV) can be measured at 10(-8) M and above. The error limits on the vanadium determinations range from +/- 3 to +/- 12% of the concentration under investigation depending on the conditions under which the assay was conducted.
Author Bunch, R L
Tawara, J
Crans, D C
Gottlieb, M S
Theisen, L A
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  givenname: L A
  surname: Theisen
  fullname: Theisen, L A
BackLink https://www.ncbi.nlm.nih.gov/pubmed/2221369$$D View this record in MEDLINE/PubMed
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Snippet A kinetic method based on alkaline phosphatase has been developed to measure free trace levels of vanadium(IV) and (V). The method involves measuring the rate...
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StartPage 53
SubjectTerms Alkaline Phosphatase - antagonists & inhibitors
Buffers
Hydrogen-Ion Concentration
Indicators and Reagents
Kinetics
Magnetic Resonance Spectroscopy
Radioisotopes
Reference Standards
Spectrophotometry, Ultraviolet
Temperature
Vanadium - analysis
Vanadium - pharmacology
Title A kinetic method for determination of free vanadium(IV) and (V) at trace level concentrations
URI https://www.ncbi.nlm.nih.gov/pubmed/2221369
Volume 188
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