Chemiluminescent compositions, enhancing reagents for chemiluminescence and methods for the preparation and use thereof

A chemiluminescent enhancing reagent configured to enhance the chemiluminescence of a 1,2-dioxetane compound having the structure:

• wherein R1 is selected from alkyl, aryl, aralkyl, alkaryl, heteroalkyl, heteroaryl, cycloalkyl, heterocycloalkyl, alkyloxyalkyl, alkyloxyaryl, alkyloxyhaloalkyl, alkylalkenyl, alkylalkynyl, halogenated alkyl, alkyl alcohol, alkyl carbonitrile, alkyl amine, alkyl acid, halogenated alkyl alcohol, halogenated alkyl carbonitrile, halogenated alkyl amine, or halogenated alkyl acid;
• X is hydrogen or a substituent on phenyl ring;
• Y is hydrogen, alkyl, acetoxyl, t-butyldimethylsilyl, a group capable of being cleaved by an enzyme, or a group capable of being cleaved by an antibody;
• and Z is hydrogen or a substituent on adamantane ring,
• wherein the chemiluminescent enhancing reagent comprises:
• at least one alkyl dimethyl benzyl quaternary ammonium salt having the structure of general formula I:
• wherein:
• R is selected from C8-20 alkyl or C8-20 alkenyl;
• X− is a negative ion;
• Y is a benzene ring substituent, each of which is independently selected from hydrogen, halogen, hydroxyl, sulfhydryl, cyano, nitro, alkyl, alkoxy, haloalkyl, amino, alkylamino, and amido;
• a fluorescent agent;
• and
• a solvent configured to dissolve the at least one alkyl dimethyl benzyl quaternary ammonium salt, the fluorescent agent, and the 1,2-dioxetane compound;
• wherein the final concentration of the alkyl dimethyl benzyl quaternary ammonium salt is in a range of between about 1 g/L to about 10 g/L;
• and
• wherein the relative final concentration of the alkyl dimethyl benzyl quaternary ammonium salt to the final concentration of the fluorescent agent is between about 1 g/L to about 2 g/L of alkyl dimethyl benzyl quaternary ammonium salt to about 1 mg/L of fluorescent agent.

A kit for enhancing the chemiluminescence of 1,2-dioxetane compounds having the structure:

• wherein R1 is selected from alkyl, aryl, aralkyl, alkaryl, heteroalkyl, heteroaryl, cycloalkyl, heterocycloalkyl, alkyloxyalkyl, alkyloxyaryl, alkyloxyhaloalkyl, alkylalkenyl, alkylalkynyl, halogenated alkyl, alkyl alcohol, alkyl carbonitrile, alkyl amine, alkyl acid, halogenated alkyl alcohol, halogenated alkyl carbonitrile, halogenated alkyl amine, or halogenated alkyl acid;
• X is hydrogen or a substituent on phenyl ring;
• Y is hydrogen, alkyl, acetoxyl, t-butyldimethylsilyl, a group capable of being cleaved by an enzyme, or a group capable of being cleaved by an antibody;
• and Z is hydrogen or a substituent on adamantane ring;
• wherein the kit comprises:
• at least one alkyl dimethyl benzyl quaternary ammonium salt having the structure of General Formula I:
• wherein:
• R is selected from C8-20 alkyl or C8-20 alkenyl;
• X− is a negative ion;
• Y is a benzene ring substituent, each of which is independently selected from:
• hydrogen, halogen, hydroxyl, sulfhydryl, cyano, nitro, alkyl, alkoxy, haloalkyl, amino, alkylamino, and amido;
• a fluorescent agent;
• and
• a manual instructing a user to mix the alkyl dimethyl benzyl quaternary ammonium salt with, the fluorescent agent and a 1,2-dioxetane compound, wherein the final concentration of the alkyl dimethyl benzyl quaternary ammonium salt is in a range of between about 1 g/L to about 10 g/L, and wherein the relative final concentration of the alkyl dimethyl benzyl quaternary ammonium salt to the final concentration of the fluorescent agent is between about 1 g/L to about 2 g/L of alkyl dimethyl benzyl quaternary ammonium salt to about 1 mg/L of fluorescent agent.

A method for enhancing the chemiluminescence of 1,2-dioxetane compounds having the structure:

• wherein R1 is selected from alkyl, aryl, aralkyl, alkaryl, heteroalkyl, heteroaryl, cycloalkyl, heterocycloalkyl, alkyloxyalkyl, alkyloxyaryl, alkyloxyhaloalkyl, alkylalkenyl, alkylalkynyl, halogenated alkyl, alkyl alcohol, alkyl carbonitrile, alkyl amine, alkyl acid, halogenated alkyl alcohol, halogenated alkyl carbonitrile, halogenated alkyl amine, or halogenated alkyl acid;
• X is hydrogen or a substituent on phenyl ring;
• Y is hydrogen, alkyl, acetoxyl, t-butyldimethylsilyl, a group capable of being cleaved by an enzyme, or a group capable of being cleaved by an antibody;
• and Z is hydrogen or a substituent on adamantane ring,
• wherein the method comprises:
• providing at least one alkyl dimethyl benzyl quaternary ammonium salt having the structure of General Formula I:
• wherein:
• R is selected from C8-20 alkyl or C8-20 alkenyl;
• X− is a negative ion;
• Y is a benzene ring substituent, each of which is independently selected from:
• hydrogen, halogen, hydroxyl, sulfhydryl, cyano, nitro, alkyl, alkoxy, haloalkyl, amino, alkylamino, and amido;
• mixing the alkyl dimethyl benzyl quaternary ammonium salt with the 1,2-dioxetane compound;
• mixing the alkyl dimethyl benzyl quaternary ammonium salt and the 1,2-dioxetane compound with a fluorescent agent;
• and
• dissolving the alkyl dimethyl benzyl quaternary ammonium salt, the fluorescent agent, and the 1,2-dioxetane compound in water, wherein the final concentration of the alkyl dimethyl benzyl quaternary ammonium salt is in a range of between about 1 g/L to about 10 g/L, and wherein the relative final concentration of the alkyl dimethyl benzyl quaternary ammonium salt to the final concentration of the fluorescent agent is between about 1 g/L to about 2 g/L of alkyl dimethyl benzyl quaternary ammonium salt to about 1 mg/L of fluorescent agent.