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

1. A chemiluminescent composition, comprising a 1,2-dioxetane compound having the structure:

wherein R 1 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;

at least one alkyl dimethyl benzyl quaternary ammonium salt having the structure of General Formula I:

wherein:

R is selected from C 8-20 alkyl or C 8-20 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;

wherein the 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

a fluorescent agent;

wherein the relative concentration of the alkyl dimethyl benzyl quaternary ammonium salt to the concentration of the fluorescent agent in the chemiluminescent composition 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.

2. The chemiluminescent composition of claim 1 , wherein R is C 12-18 alkyl.

3. The chemiluminescent composition of claim 1 , wherein X − is at least one of the following: a halide ion, ClO 4 − , PF 6 − , CF 3 SO 3 − , BF 4 − , acetate, and p-toluene sulfonate.

4. The chemiluminescent composition of claim 1 , wherein the alkyl dimethyl benzyl quaternary ammonium salt is selected from at least one of the following: a hexadecyl dimethyl benzyl ammonium salt, an octadecyl dimethyl benzyl ammonium salt, a dodecyl dimethyl benzyl ammonium salt, a myristyl dimethyl benzyl ammonium salt and combinations thereof.

5. The chemiluminescent composition of claim 1 , wherein the alkyl dimethyl benzyl quaternary ammonium salt is selected from at least one of the following: hexadecyl dimethyl benzyl ammonium chloride, octadecyl dimethyl benzyl ammonium chloride, dodecyl dimethyl benzyl ammonium chloride, myristyl dimethyl benzyl ammonium chloride, dodecyl dimethyl benzyl ammonium bromide, myristyl dimethyl benzyl ammonium bromide, hexadecyl dimethyl benzyl ammonium bromide, octadecyl dimethyl benzyl ammonium bromide and combinations thereof.

6. The chemiluminescent composition of claim 1 , wherein the chemiluminescent composition further comprises at least one of: 2-amino-2-methyl-1-propanol or a magnesium ion or both.

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

wherein R 1 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 C 8-20 alkyl or C 8-20 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.

8. The enhancing reagent of claim 7 , wherein R is C 12-18 alkyl.

9. The enhancing reagent of claim 7 , wherein the alkyl dimethyl benzyl quaternary ammonium salt is selected from at least one of the following: a hexadecyl dimethyl benzyl ammonium salt, an octadecyl dimethyl benzyl ammonium salt, a dodecyl dimethyl benzyl ammonium salt, a myristyl dimethyl benzyl ammonium salt and combinations thereof.

10. The enhancing reagent of claim 7 , wherein the alkyl dimethyl benzyl quaternary ammonium salt is selected from at least one of the following: hexadecyl dimethyl benzyl ammonium chloride, octadecyl dimethyl benzyl ammonium chloride, dodecyl dimethyl benzyl ammonium chloride, myristyl dimethyl benzyl ammonium chloride, dodecyl dimethyl benzyl ammonium bromide, myristyl dimethyl benzyl ammonium bromide, hexadecyl dimethyl benzyl ammonium bromide, octadecyl dimethyl benzyl ammonium bromide and combinations thereof.

11. The enhancing reagent of claim 7 , wherein the enhancing reagent further comprises at least one of: 2-amino-2-methyl-1-propanol or a magnesium ion or both.

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

wherein R 1 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 C 8-20 alkyl or C 8-20 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.

13. The kit of claim 12 , wherein the components of the kit are stored individually or in the form of one or more mixtures.

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

wherein R 1 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 C 8-20 alkyl or C 8-20 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.

15. The method of claim 14 , wherein R is C 12-18 alkyl.

16. The method of claim 14 , wherein the alkyl dimethyl benzyl quaternary ammonium salt is selected from at least one of the following: hexadecyl dimethyl benzyl ammonium chloride, octadecyl dimethyl benzyl ammonium chloride, dodecyl dimethyl benzyl ammonium chloride, myristyl dimethyl benzyl ammonium chloride, dodecyl dimethyl benzyl ammonium bromide, myristyl dimethyl benzyl ammonium bromide, hexadecyl dimethyl benzyl ammonium bromide, octadecyl dimethyl benzyl ammonium bromide and combinations thereof.

17. The method of claim 14 , further comprising mixing the alkyl dimethyl benzyl quaternary ammonium salt and the 1,2-dioxetane compound with at least one of: 2-amino-2-methyl-1-propanol or a magnesium ion or both.