Method and compositions providing enhanced chemiluminescence from 1,2-dioxetanes

Claim 2

A composition which generates light upon triggering which comprises in admixture: (a) a fluorescent compound; and (b) a stable 1,2-dioxetane of the formula: ##STR32## wherein ArOX represents an aryl group substituted with an X-oxy group which forms an unstable oxide intermediate 1,2-dioxetane compound when triggered to remove X by an activating agent so that the unstable 1,2-dioxetane compound decomposes and releases electronic energy to form light and two carbonyl containing compounds of the formula: ##STR33## wherein X is a chemically labile group which is removed by the activating agent to form the unstable oxide intermediate 1,2-dioxetane and wherein A are passive organic groups which allow the light to be produced wherein the stable 1,2-dioxetane is decomposed with the activating agent wherein the dioxetane and the fluorescent compound are provided in a closely spaced relationship in the presence of a surfactant, micelle, liposome, reversed micelle, microemulsion, film, including a monolayer, or polymer and wherein the fluorescent compound accepts the electronic energy generated upon decomposition of the unstable oxide intermediate and produces a more intense light than is produced by the triggering of the dioxetane alone.

Claim 3

A composition which generates light upon triggering which comprises in admixture: (a) a fluorescent compound; and (b) a stable 1,2-dioxetane compound of the formula: ##STR34## wherein one of R 1 and R 2 together and R 3 and R 4 together can be joined as spirofused alkylene which can contain hetero atoms selected from the group consisting of N, S, O and P in addition to carbon and aryl rings, wherein at least one of R 1 and R 2 or R 3 and R 4 is an aryl group substituted with an X-oxy group which forms an unstable oxide intermediate 1,2 dioxetane compound when triggered to remove X by an activating agent selected from acids, bases, salts, enzymes, inorganic and organic catalysts and electron donors so that the unstable 1,2-dioxetane compound decomposes and releases electronic energy to form light and two carbonyl containing compounds of the formula: ##STR35## wherein those of R 1 , R 2 , R 3 or R 4 which are unsubstituted by an X-oxy group are organic groups which provide stability for the stable 1,2-dioxetane compound and wherein X is a chemically labile group which is removed by the activating agent to form the unstable oxide intermediate wherein the stable 1,2-dioxetane is decomposed with the activating agent, wherein the dioxetane and the fluorescent compound are provided n a closely spaced relationship in the presence of a surfactant, micelle, liposome, reversed micelle, microemulsion, film, including a monolayer, or polymer and wherein the fluorescent compound accepts the electronic energy generated upon decomposition of the unstable oxide intermediate and produces a more intense light than is produced by the triggering of the dioxetane alone.

Claim 4

A composition which generates light upon triggering which comprises in admixture: (a) a fluorescent compound; and (b) a stable 1,2-dioxetane compound of the formula: ##STR34## wherein one of R 1 and R 2 together and R 3 and R 4 together can be joined as spirofused alkylene which can contain hetero atoms selected from the group consisting of N, S, O and P in addition to carbon and aryl rings, wherein at least one of R 1 and R 2 or R 3 and R 4 is an aryl group substituted with an X-oxy group which forms an unstable oxide intermediate 1,2 dioxetane compound when triggered to remove X by an activating agent selected from acids, bases, salts, enzymes, inorganic and organic catalysts and electron donors so that the unstable 1,2-dioxetane compound decomposes and releases electronic energy to form light and two carbonyl containing compounds of the formula: ##STR35## wherein those of R 1 , R 2 , R 3 or R 4 which are unsubstituted by an X-oxy group are organic groups which provide stability for the stable 1,2-dioxetane compound and wherein X is a chemically labile group which is removed by the activating agent to form the unstable oxide intermediate wherein the stable 1,2-dioxetane is decomposed with the activating agent, wherein the dioxetane and the fluorescent compound are provided n a closely spaced relationship in the presence of a surfactant, micelle, liposome, reversed micelle, microemulsion, film, including a monolayer, or polymer and wherein the fluorescent compound accepts the electronic energy generated upon decomposition of the unstable oxide intermediate and produces a more intense light than is produced by the triggering of the dioxetane alone.

Claim 5

A composition which generates light upon triggering which comprises in admixture: (a) a fluorescent compound; and (b) a stable dioxetane compound of the formula: ##STR36## wherein R 1 is selected from alkyl, alkoxy, aryloxy dialkylamino, trialkyl or aryl silyloxy and aryl groups including spirofused aryl groups with R 2 , wherein R 2 si an aryl group which can include R 1 and is substituted with an X-oxy group which forms an unstable oxide intermediate 1,2-dioxetane compound when triggered to remove X by an activating agent selected from acids, bases, salts, enzymes, inorganic and organic catalysts and electron donors so that the unstable 1,2-dioxetane compound decomposes and releases electronic energy to form light and two carbonyl containing compounds of the formula: ##STR37## wherein X is a chemically labile group which is removed by the activating agent to form the unstable oxide intermediate and wherein R 3 and R 4 are selected from aryl, heteroalkyl and alkyl groups which can be joined together as spirofused polycyclic alkyl and polycyclic aryl groups wherein the stable 1,2-dioxetane is decomposed with the activating agent, wherein the dioxetane and the fluorescent compound are provided in a closely spaced relationship in the presence of a surfactant, micelle, liposome, reversed micelle, microemulsion, film, including a monolayer, or polymer and wherein the fluorescent compound accepts the electronic energy generated upon decomposition of the unstable oxide intermediate and produces a more intense light than is produced by the triggering of the dioxetane alone.

Claim 6

A composition which generates light upon triggering which comprises in admixture: (a) a fluorescent compound; and (b) a stable dioxetane compound of the formula: ##STR36## wherein R 1 is selected from alkyl, alkoxy, aryloxy dialkylamino, trialkyl or aryl silyloxy and aryl groups including spirofused aryl groups with R 2 , wherein R 2 si an aryl group which can include R 1 and is substituted with an X-oxy group which forms an unstable oxide intermediate 1,2-dioxetane compound when triggered to remove X by an activating agent selected from acids, bases, salts, enzymes, inorganic and organic catalysts and electron donors so that the unstable 1,2-dioxetane compound decomposes and releases electronic energy to form light and two carbonyl containing compounds of the formula: ##STR37## wherein X is a chemically labile group which is removed by the activating agent to form the unstable oxide intermediate and wherein R 3 and R 4 are selected from aryl, heteroalkyl and alkyl groups which can be joined together as spirofused polycyclic alkyl and polycyclic aryl groups wherein the stable 1,2-dioxetane is decomposed with the activating agent, wherein the dioxetane and the fluorescent compound are provided in a closely spaced relationship in the presence of a surfactant, micelle, liposome, reversed micelle, microemulsion, film, including a monolayer, or polymer and wherein the fluorescent compound accepts the electronic energy generated upon decomposition of the unstable oxide intermediate and produces a more intense light than is produced by the triggering of the dioxetane alone.

Claim 7

The composition of claim 1 wherein the mole ratio of dioxetane to fluorescent compound is between about 1,000 to 1 and 1 to 1.

Claim 8

The composition of claim 1 wherein the fluorescent compound is a fluorescein compound.

Claim 9

The composition of claim 1 wherein in addition a surfactant is provided in the composition.

Claim 10

The composition of claim 2 wherein R 1 is a methoxy group and R 2 is a phenyl group substituted with an X-oxy group and wherein R 3 and R 4 are joined together as an adamantyl group.

Claim 11

The composition of claim 7 wherein the X-oxy group is selected from hydroxyl, trialkyl or aryl silyloxy, inorganic oxyacid salt, oxygen-pyranoside, arylcarboxyl esters and alkylcarboxyl esters and wherein X is removed by the activating agent.

Claim 12

The composition of claim 8 wherein the fluorescent compound is a fluorescein compound.

Claim 13

The composition of claim 1 wherein a cationic surfactant is provided in the composition, wherein the fluorescent compound is chemically linked with a hydrocarbon chain to provide a fluorescent hydrocarbon which acts as a co-surfactant with a cationic surfactant to increase the light from the dioxetane and fluorescent compound and wherein the dioxetane is present in an amount between about 10 -2 to 10 -6 M, the surfactant is present in an amount greater than 10 -4 M and the co-surfactant is present in an amount between 10 -3 to 10 -6 M.

Claim 14

The composition of claim 10 wherein the cationic surfactant is a cetyltrimethylammonium salt.

Claim 15

The composition of claim 1 wherein the dioxetane and fluorescent compound are in a closely spaced relationship in a micelle, liposome, reversed micelle, microemulsion, film, monolayer or polymer.

Claim 16

The composition of claim 1 wherein the 1,2-dioxetane has the formula: ##STR38## ps wherein X is a group reactive with the activating agent to produce the two carbonyl containing compounds ##STR39## and light.

Claim 17

The composition of claim 13 wherein OX is a phosphate group and the activating agent is alkaline phosphatase.

Claim 18

The composition of claim 13 wherein in addition a surfactant is admixed in the composition.

Claim 19

The composition of claim 15 wherein the surfactant is a cetyltrimethylammonium salt.

Claim 20

The composition of claim 13 wherein the fluorescent compound is a fluorescein compound.

Claim 21

The composition of claim 17 wherein in addition a surfactant is admixed in the composition.

Claim 22

The composition of claim 18 wherein the surfactant is a cetyltrimethylammonium salt.

Claim 23

The composition of claim 17 wherein the fluorescein compound is fluorescein chemically linked to a hydrocarbon to provide a fluorescein surfactant.

Claim 24

The composition of claim 13 wherein the fluorescent compound is selected from the group consisting of 5-N-tetradecanoylaminofluorescein and 1-hexadecyl-6-hydroxy-2-benzothiazamide.

Claim 25

The composition of claim 21 wherein in addition a non-fluorescent surfactant is admixed in the composition.

Claim 26

The composition of claim 22 wherein the surfactant is a cetyltrimethylammonium salt.

Claim 27

The composition of claim 1 wherein the fluorescent compound is fluorescein chemically linked to a hydrocarbon to provide a fluorescein surfactant which increases the light from the fluorescein compound and the dioxetane.

Claim 28

The composition of claim 1 wherein the fluorescent compound is selected from the group consisting of 5-N-tetradecanoylaminofluorescein and 1-hexadecyl-6-hydroxy-2-benzothiazamide.

Claim 29

The composition of claim 25 wherein in addition a surfactant is admixed in the composition.

Claim 30

The composition of claim 26 wherein the surfactant is a cetyltrimethylammonium salt.

Claim 31

The composition of claim 3 wherein the fluorescent compound is fluorescein chemically linked to a hydrocarbon to provide a fluorescein surfactant.

Claim 32

The composition of claim 3 wherein the fluorescent compound is selected from the group consisting of 5-N-tetradecanoylaminofluorescein and 1-hexadecyl-6-hydroxy-2-benzothiazamide.

Claim 33

The composition of claim 29 wherein in addition a surfactant is admixed in the composition.

Claim 34

The composition of claim 30 wherein the surfactant is a cetyltrimethylammonium salt.

Claim 35

The composition of claim 1 wherein the dioxetane and the fluorescent compound are provided in a closely spaced relationship in a solid or liquid medium.

Claim 36

The composition of claim 2 wherein the dioxetane and the fluorescent compound are provided in a closely spaced relationship in a solid or liquid medium.

Claim 37

The composition of claim 3 wherein the dioxetane and the fluorescent compound are provided in a closely spaced relationship in a solid or liquid medium.

Claim 38

The composition of claim 3 wherein in addition a surfactant is provided in the composition.