Method for the determination of the concentration of vitamin b6 in a sample
AssigneeBuehlmann Laboratories AG
InventorsJakob Weber, Lara Hasan, Thomas Jermann, Michel-Angelo Sciotti, Daniel Gygax, Andre Scholer
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Abstract
The invention relates to methods for the determination of vitamin B6 in samples as well as to reagent compositions for assaying a sample for vitamin B6 and to a test kit suitable for carrying out the methods according to the present invention. Further, the invention relates to the use of such methods for the application to different analyzing devices such as micro-titer plate readers and fully automated clinical chemistry analyzers (autoanalyzers).
Claims25
A method to determine the concentration of pyridoxal-5′-phosphate (PLP, vitamin B6) in a sample, the method comprising the steps of
- a) incubation of the sample containing PLP with at least one PLP-dependent enzyme,
- b) contacting the PLP-dependent enzyme(s) from step a) with tyrosine,
- c) enzymatic conversion of the tyrosine to tyramine by a decarboxylase,
- d) enzymatic conversion of the tyramine resulting from step c) to hydrogen peroxide and at least one other pro-duct by an oxidase,
- e) enzymatic reaction of at least one indicator substrate and the hydrogen peroxide of step d) to a detectable compound by a peroxidase,
- f) measuring the quantity of the detectable compound of step e)
- and
- g) correlating the measured quantity of step f) with the concentration of pyridoxal-5′-phosphate (PLP, vitamin B6) in the sample.
A composition for assaying a sample for pyridoxal-5′-phosphate (PLP, vitamin B6), comprising:
- a) a decarboxylase capable of the conversion of tyrosine to tyramine,
- b) tyrosine,
- c) an oxidase capable of the conversion of tyramine to hydrogen peroxide and at least one other product,
- d) a peroxidase capable of the conversion of at least one indicator substrate to a detectable compound in the presence of hydrogen peroxide.
Dependencies are inferred from the claim text.