Aryl N-alkylacridanthiocarboxylate derivatives useful for chemiluminescent detection

A reagent composition which generates light in the presence of a peroxidase comprising

• a) an acridan of the formula:
• ##STR17## wherein R is selected from the group consisting of alkyl, heteroalkyl and aralkyl groups, wherein R.sub.1 through R.sub.8 are independently selected from the group consisting of hydrogen and groups which do not interfere with the production of light and wherein Ar is selected from the group consisting of substituted and unsubstituted aryl groups which do not interfere with the production of light from the acridan by reaction with a peroxide and a peroxidase;
• b) optionally a phenolic compound which enhances light production;
• c) the peroxide which participates in the reaction of the acridan with the peroxidase;
• d) a chelating agent which prevents the peroxide compound from reacting prior to addition of the peroxidase to the composition;
• and
• e) a surfactant in an amount which provides improved chemiluminescence by reducing background.

A method of detecting an analyte selected from the group consisting of peroxidase alone, organic molecules labeled with the peroxidase and biological molecules labeled with the peroxidase in an assay procedure by a chemiluminescent reaction comprising:

• a) providing a reagent composition which generates light in the presence of a peroxidase which comprises an acridan of the formula:
• ##STR20## wherein R is selected from the group consisting of alkyl, heteroalkyl and aralkyl groups, wherein R.sub.1 through R.sub.8 are independently selected from the group consisting of hydrogen and groups which do not interfere with the production of light and wherein Ar is selected from the group consisting of substituted and unsubstituted aryl groups which do not interfere with the production of light from the acridan by reaction with a peroxide and a peroxidase;
• optionally a phenolic compound which enhances light production from the acridan;
• the peroxide which participates in the reaction of the acridan with the peroxidase;
• a chelating agent which prevents the peroxide from reacting prior to addition of the peroxidase to the composition;
• and a surfactant in an amount which provides improved chemiluminescene by reducing background;
• and
• b) reacting the peroxidase with the reagent composition so that light is produced to detect the analyte.

A kit for detecting an analyte selected from the group consisting of hydrogen peroxide, hydrogen peroxide generated by an enzyme, peroxidase alone, organic molecules labeled with the peroxidase and biological molecules labeled with the peroxidase in an assay procedure by a chemiluminescent reaction comprising in separate containers:

• a) an acridan of the formula:
• ##STR21## wherein R is selected from the group consisting of alkyl, heteroalkyl and aralkyl groups, wherein R.sub.1 through R.sub.8 are independently selected from the group consisting of hydrogen and groups which do not interfere with the production of light and wherein Ar is selected from the group consisting of substituted and unsubstituted aryl groups which do not interfere with the production of light from the acridan by reaction with a peroxide and the peroxidase;
• b) the peroxide and
• c) the peroxidase and enzyme either singly or attached to an analyte-binding compound, wherein the light is detected in the assay procedure by reacting the acridan compound with the peroxide and the peroxidase enzyme to detect the analyte.

The kit for detecting an analyte selected from the group consisting of peroxidase alone, organic molecules labeled with the peroxidase and biological molecules labeled with the peroxidase in an assay procedure by a chemiluminescent reaction comprising providing in separate containers:

• a) a reagent composition which generates light in the presence of the peroxidase which comprises an acridan of the formula:
• ##STR22## wherein R is selected from the group consisting of alkyl, heteroalkyl and aralkyl groups, wherein R.sub.1 through R.sub.8 are independently selected from the group consisting of hydrogen and groups which do not interfere with the production of light and wherein Ar selected from the group consisting of substituted and unsubstituted aryl groups which do not interfere with the production of light from the acridan by reaction with a peroxide an the peroxidase;
• optionally a phenolic compound which enhances light production from the acridan;
• the peroxide which participates in the reaction of the acridan with the peroxidase;
• a chelating agent which prevents the peroxide compound from reacting prior to addition of the peroxidase to the composition;
• and a surfactant in an amount which provides improved chemiluminescene which reduces background;
• and
• b) the peroxidase enzyme either singly or attached to an analyte-binding compound, wherein the light is detected in the assay procedure by reacting the reagent composition with the peroxidase enzyme to detect the analyte.