Biochemical method to measure niacin status in a biological sample
AssigneeNiadyne INC
InventorsElaine L. Jacobson, Myron K. Jacobson
Life of this patent
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Abstract
The present invention is a novel and rapid assay for intracellular niacin status in a biological sample that is both highly sensitive and accurate over a wide range of niacin status. The assay can be performed directly on a biological sample such as blood and is useful for determining the niacin status in a subject such as a human. In a preferred form, the assay is incorporated into a kit or an apparatus for the rapid and accurate determination of niacin status in a subject.
Claims31
A method of assaying an intracellular pyridine nucleotide status of a biological sample comprising:
- a) obtaining the biological sample;
- b) extracting the intracellular pyridine nucleotide from the biological sample;
- c) subjecting said pyridine nucleotide in the sample to a cycling process comprising one or more cycles of:
- reducing the pyridine nucleotide to a reduced pyridine nucleotide;
- transferring an electron from the reduced pyridine nucleotide to an electron acceptor dye molecule to produce a change in an absorbance of the electron acceptor dye molecule;
- and
- d) measuring the change in absorbance of the electron acceptor dye molecule to determine the intracellular pyridine nucleotide status.
A method to measure intracellular niacin status in whole blood comprising:
- a) collecting a whole blood sample;
- b) extracting intracellular pyridine nucleotide from the whole blood sample;
- c) subjecting said pyridine nucleotide in the sample to a cycling process comprising one or more cycles of:
- reducing the pyridine nucleotide to a reduced pyridine nucleotide;
- transferring an electron from the reduced pyridine nucleotide to an electron acceptor dye molecule to produce a change in an absorbance of the electron acceptor dye molecule;
- d) measuring the change in absorbance of the electron acceptor dye molecule to determine the intracellular pyridine nucleotide status.
A method for detecting the amount of a reduced intracellular pyridine nucleotide in a biological sample comprising
- a) obtaining the biological sample;
- b) extracting the reduced intracellular pyridine nucleotide from said sample with hot alkaline treatment;
- c) neutralizing said sample;
- d) subjecting said reduced pyridine nucleotide in the sample to a cycling process comprising one or more cycles of:
- reducing the pyridine nucleotide to a reduced pyridine nucleotide;
- transferring an electron from the reduced pyridine nucleotide to an electron acceptor dye molecule to produce a change in an absorbance of the electron acceptor dye molecule;
- and
- e) measuring the change in absorbance of the electron acceptor dye molecule to determine the intracellular reduced pyridine nucleotide.
A method for detecting the amount of total intracellular pyridine nucleotide in a biological sample comprising
- a) obtaining the biological sample from a subject;
- b) extracting the total intracellular pyridine nucleotide from said sample with cold alkaline treatment;
- c) neutralizing said sample;
- d) subjecting the total pyridine nucleotide in the sample to a cycling process comprising one or more cycles of:
- reducing the pyridine nucleotide to a reduced pyridine nucleotide;
- transferring an electron from the reduced pyridine nucleotide to an electron acceptor dye molecule to produce a change in an absorbance of the electron acceptor dye molecule;
- and
- e) measuring the change in absorbance of the electron acceptor dye molecule to determine the total intracellular reduced pyridine nucleotide.
A kit for the extraction of pyridine nucleotides from a biological sample comprising:
- a base solution;
- an acid solution;
- one or more neutralizing solutions to neutralize the base solution and the acid solution;
- and
- an electron acceptor dye.
Dependencies are inferred from the claim text.