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

1. A method for enhancing the chemiluminescence of 1,2-dioxetane compounds, the method comprising:

providing at least one alkyl bis-quaternary ammonium salt having the structure of General Formula I:

wherein:

R 1 is selected from at least one of the following: C 10-20 alkyl, C 10-20 alkenyl, C 10-20 alkynyl, and C 3-8 cycloalkylC 6-12 alkyl-;

each n is an integer independently selected from 2 to 15;

each m is an integer independently selected from 1 to 6;

R 2 , R 3 , R 4 , R 5 , R 6 , and R 7 are each independently selected from at least one of the following: C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, and C 3-8 cycloalkyl; and

X − is a negative ion; and

mixing the alkyl bis-quaternary ammonium salt with a 1,2-dioxetane compound.

2. The method of claim 1 , wherein R 1 is selected from C 12-18 alkyl.

3. The method of claim 1 , wherein R 2 , R 3 , R 4 , R 5 , R 6 , and R 7 are each independently selected from methyl or ethyl.

4. The method of claim 1 , wherein R 2 , R 3 , R 4 , R 5 , R 6 , and R 7 are alkyl, each having the same number of carbon atoms.

5. The method of claim 1 , wherein each n is an integer independently selected from 5 to 10, or each m is independently selected from 2 or 3.

6. The method of claim 1 , wherein X − is selected from at least one of the following: halide ions, ClO 4 − , PF 6 − , CF 3 SO 3 − , BE 4 − , acetate, and p-toluene sulfonate anions.

7. The method of claim 1 , wherein the alkyl bis-quaternary ammonium salt compound is selected from at least one of the following: dodecylamino polyoxyethylene ether ammonium dichloride, octadecylamino polyoxyethylene ether ammonium dichloride, dodecylamino polyoxyethylene ether ammonium dibromide, octadecylamino polyoxyethylene ether ammonium dibromide, and a combination thereof.

8. The method of claim 1 , further comprising dissolving the alkyl bis-quaternary ammonium salt and the 1,2-dioxetane compound in water, wherein the concentration of the alkyl bis-quaternary ammonium salt is in a range of between about 0.1 g/L and about 10 g/L.

9. The method of claim 1 , further comprising dissolving the alkyl bis-quaternary ammonium salt and the 1,2-dioxetane compound in water, wherein the concentration of the alkyl bis-quaternary ammonium salt is in a range of between about 1 g/L and about 10 g/L.

10. The method of claim 1 , further comprising mixing the alkyl bis-quaternary ammonium salt and the 1,2-dioxetane compound with a fluorescent agent.

11. The method of claim 1 , further comprising mixing the alkyl bis-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.

12. A method of preparing an enhancing reagent for chemiluminescence of 1,2-dioxetane compounds, comprising:

providing an alkyl bis-quaternary ammonium salt having the structure of General Formula I:

wherein:

R 1 is selected from at least one of the following: C 10-20 alkyl, C 10-20 alkenyl, C 10-20 alkynyl, and C 3-8 cycloalkylC 6-12 alkyl;

each n is an integer independently selected from 2 to 15;

each m is an integer independently selected from 1 to 6;

R 2 , R 3 , R 4 , R 5 , R 6 , and R 7 are each independently selected from at least one of the following: C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, and C 3-8 cycloalkyl; and

X − is a negative ion;

adding, to purified water, the alkyl bis-quaternary ammonium salt having the structure of General Formula I at a concentration ranging from about 0.1 g/L to about 10 g/L;

adding a 1,2-dioxetane compound at a concentration ranging from about 0.0001 mol/L to about 0.01 mol/L;

optionally adding a fluorescent agent to a concentration ranging from about 0.1 mg/L to about 1 g/L; and

optionally adding magnesium ions to a concentration ranging from about 0.001 mol/L to about 0.01 mol/L and

optionally adding a buffering agent to a concentration ranging from about 10 mM to about 500 mM; and

mixing the added components in the purified water to form an enhancing reagent for chemiluminescence of 1,2-dioxetane compounds.