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US 5,043,269 AGranted
Chromogenic substrate to peroxidase enzymes
OwnerSpyros Theodoropulosinventor
Life of this patent
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Abstract
A novel chromogenic substrate to peroxidase enzymes is provided which is comprised of a mixture of a colorless mixture of two compounds, such as, 3-methyl-2-benzothiazolinone hydrazone and 2-hydroxy-2,4,6-cycloheptatrienone, which undergo an oxidative coupling in the presence of peroxidase and hydrogen peroxide forming a purple indamine dye. Under certain circumstances the dye will precipitate out from the buffered solution and thus provide a permanent record of the determination. The mixture is also stable and unaffected by oxygen of the air or by hydrogen peroxide.
Claims10
A chromogenic substrate for peroxidase comprising a mixture of:
- (a) a hydrazone compound of the formula:
- ##STR6## and (b) a 2-hydroxy-2,4,6-cycloheptatrienone compound of the formula:
- ##STR7## wherein:
- R represents a group containing from 1 to 25 carbon atoms and is selected from the group consisting of alkyl, alkenyl, aralkyl, hydroxyalkyl, alkoxyalkyl, aryloxyalkyl, carboxyalkyl, carboalkoxyalkyl, arylthioalkyl, suflonalkyl;
- R.sub.1 -R.sub.4 represent hydrogen, halogen, NO, NO.sub.2, SO.sub.3 H, COOH, or a group containing from 1 to 25 carbon atoms which can be substituted or unsubstituted alkyl, alkoxy, aryl, heteroaryl or aryloxy;
- X represents an anion selected from the group consisting of F, Cl, Br, I, acetate, phthalate, phosphate, citrate, borate, sulfate and nitrate;
- and
- Z represents a group containing non-metallic atoms necessary to complete a heterocyclic or heterobicyclic ring with the atoms to which it is attached and contains at least one member selected from the group consisting of oxygen and sulfur and up to a total of 25 carbon atoms, and can be substituted with one or more substituents selected from the group consisting of lower alkyl, nitro, halogen, carboxyl, sulfonyl, amino and diamino groups.
5 more elements
Dependencies are inferred from the claim text.