Method, composition and elements for the detecting of nitrogen-c

Details Method, composition and elements for the detecting of nitrogen-c Method, composition and elements for the detecting of nitrogen-c

1978-02-24

1980-03-18

Shapiro, Lionel M.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving urea or urease

435 4, G01N 3314

Patent

active

041940636

ABSTRACT:
Methods, compositions and elements for detecting a nitrogen-containing compound capable of releasing ammonia upon enzymatic action are described. The method comprises contacting in an aqueous medium a sample suspected of containing such a compound and a novel assay composition comprising at least one enzyme which catalyzes the decomposition of the compound to ammonia and detecting the ammonia by condensation with a .beta.-diketone in the presence of an aldehyde, preferably formaldehyde, and measuring the color or fluorescence produced by any resulting dihydropyridine. According to a preferred embodiment, the aldehyde is generated in situ and the .beta.-diketone is polymeric.
The foregoing assay composition can be incorporated into single-layer or multilayer analytical elements of the type known in the prior art. A preferred such element comprises an isotropically porous spreading layer in fluid contact with a reagent layer. The spreading layer is most preferably non-fibrous.

REFERENCES:
patent: Re27524 (1972-11-01), Nadeau
patent: 3145086 (1964-08-01), Free
patent: 3806416 (1974-04-01), Mollering et al.
patent: 3873269 (1975-03-01), Kraffczyk et al.
patent: 3992158 (1976-11-01), Przybylowicz et al.
patent: 4125377 (1978-11-01), Gindler
Hantzsch, Ann. Chem. Liebigs, 215, p. 1, 1882 (translation of pp. 5-8, only).
Knoevenagel, Annalen, 281, p. 25, 1894 (translation of pp. 25-28, only).
Dunsbach in Chemical Abstracts, vol. 66 (1967) 82993a.

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