Biological and analytical uses of phenalenone and benzphenalenone compounds

Claim 2

An analytical test composition buffered to a pH of 9 or less, said composition comprising a fluorescent dye selected from the group consisting of ##STR30## wherein R is hydroxy, mercapto or amino or salts thereof.

Claim 3

The composition of claim 1 wherein amino is HN(R')-- and wherein R' is hydrogen, alkyl, phenyl, cyclohexyl or a heterocyclic group.

Claim 4

The composition of claim 1 hwerein said fluorescent dye is ##STR31## wherein R is hydroxy or amino or salts thereof.

Claim 5

An analytical composition comprising a reducible compound of the structure CAR--R 1 ) n wherein CAR-- is an aromatic or quinone nucleus having a hydrogen atom removed to provide a valence through which R 1 is attached to CAR--, R 1 comprises a fluorescent moiety derived from a compound selected from the group consisting of ##STR32## wherein R is hydroxy or mercapto or salts thereof, and n is 1 or 2, said fluorescent moiety having an oxy or thio linkage, derived from said R group by removal of a hydrogen atom, through which said moiety is attached to said CAR-- nucleus, provided CAR--R 1 ) n is constructed so that it is capable of being reduced at a pH of 9 or less to release said derived fluorescent moiety, and further provided that when R 1 is replaced with H, CAR--H) n has an E 1/2 of at least about +100 mV when measured in water.

Claim 6

An analytical composition comprising a reducible compound of the structure CAR--R 1 ) n wherein CAR-- is an aromatic or quinone nucleus having a hydrogen atom removed to provide a valence through which R 1 is attached to CAR--, R 1 comprises a fluorescent moiety derived from a compound selected from the group consisting of ##STR32## wherein R is hydroxy or mercapto or salts thereof, and n is 1 or 2, said fluorescent moiety having an oxy or thio linkage, derived from said R group by removal of a hydrogen atom, through which said moiety is attached to said CAR-- nucleus, provided CAR--R 1 ) n is constructed so that it is capable of being reduced at a pH of 9 or less to release said derived fluorescent moiety, and further provided that when R 1 is replaced with H, CAR--H) n has an E 1/2 of at least about +100 mV when measured in water.

Claim 7

The composition of claim 4 wherein said reducible compound has the structure ##STR33## wherein R 2 and R 4 are independently hydrogen, alkyl, aryl or an electron withdrawing group having a positive Hammett sigma value, R 3 is R 1 , hydrogen, alkyl, aryl or an electron withdrawing group having a positive Hammett sigma value, or R 3 and R 4 , taken together, represent the atoms necessary to complete a fused carbocyclic ring, R 5 is alkylene of 1 or 2 carbon atoms, R 6 is methy, Q is carbonyl or thiocarbonyl, FRAG is said fluorescent moiety attached to Q through said oxy or thio linkage, and m is 0 or 1.

Claim 8

The composition of claim 4 further comprising a surfactant.

Claim 9

The composition of claim 4 further comprising an electron transfer agent.

Claim 10

A dry analytical element for the determination of an analyte comprising an absorbent carrier material, and containing a reducible compound of the structure CAR--R 1 ) n wherein CAR-- is an aromatic or quinone nucleus having a hydrogen atom removed to provide a valence through which R 1 is attached to CAR--, R 1 comprises a fluorescent moiety derived from a compound selected from the group consisting of ##STR34## wherein R is hydroxyl or mercapto or salts thereof, and n is 1 or 2, said fluorescent moiety having an oxy or thio linkage, derived from said R group by removal of a hydrogen atom, through which said moiety is attached to said CAR-- nucleus, provided CAR--R 1 ) n is constructed so that it is capable of being reduced at a pH of 9 or less to release said derived fluorescent moiety, and further provided that when R 1 is replaced with H, CAR--H) n has an E 1/2 of at least about +100 mV when measured in water.

Claim 11

A dry analytical element for the determination of an analyte comprising an absorbent carrier material, and containing a reducible compound of the structure CAR--R 1 ) n wherein CAR-- is an aromatic or quinone nucleus having a hydrogen atom removed to provide a valence through which R 1 is attached to CAR--, R 1 comprises a fluorescent moiety derived from a compound selected from the group consisting of ##STR34## wherein R is hydroxyl or mercapto or salts thereof, and n is 1 or 2, said fluorescent moiety having an oxy or thio linkage, derived from said R group by removal of a hydrogen atom, through which said moiety is attached to said CAR-- nucleus, provided CAR--R 1 ) n is constructed so that it is capable of being reduced at a pH of 9 or less to release said derived fluorescent moiety, and further provided that when R 1 is replaced with H, CAR--H) n has an E 1/2 of at least about +100 mV when measured in water.

Claim 12

The element of claim 8 further containing an electron transfer agent.

Claim 13

The element of claim 8 wherein said reducible compound has the structure ##STR35## wherein R 2 and R 4 are independently hydrogen, alkyl, aryl or an electron withdrawing group having a positive Hammett sigma value, R 3 is R 1 , hydrogen, alkyl, aryl or an electron withdrawing group having a positive Hammett sigma value, or R 3 and R 4 , taken together, represent the atoms necessary to complete a fused carbocyclic ring, R 5 is alkylene of 1 or 2 carbon atoms, R 6 is methyl, Q is carbonyl or thiocarbonyl, FRAG is said fluorescent moiety attached to Q through said oxy or thio linkage, and m is 0 or 1.

Claim 14

The element of claim 10 wherein R 3 is R 1 .

Claim 15

The element of claim 10 wherein R 5 is methylene, R 6 is methyl and Q is carbonyl.

Claim 16

The element of claim 10 comprising a support carrying said absorbent carrier material.

Claim 17

A dry analytical element for making a determination of living organisms comprising a support having thereon a porous spreading zone, and containing an electron transfer agent and a reducible compound of the structure ##STR36## wherein R 2 and R 4 are independently hydrogen, alkyl, aryl or an electron withdrawing group having a positive Hammett sigma value, R 3 is R 1 , hydrogen, alkyl, aryl or an electron withdrawing group having a positive Hammett sigma value, or R 3 and R 4 , taken together, represent the atoms necessary to complete a fused carbocyclic ring, R 5 is alkylene of 1 or 2 carbon atoms, R 6 is methyl, Q is carbonyl or thiocarbonyl, FRAG is a fluorescent moiety derived from a compound selected from the group consisting of ##STR37## wherein R is hydroxy or mercapto or salts thereof, and m is 0 or 1, said fluorescent moiety having an oxy or thio linkage, derived from said R group by removal of a hydrogen atom, through which said moiety is attached to said CAR-- nucleus, provided that when R 1 is replaced with H, CAR--H has an E 1/2 of at least about +100 mV when measured in water.

Claim 18

A dry analytical element for making a determination of living organisms comprising a support having thereon a porous spreading zone, and containing an electron transfer agent and a reducible compound of the structure ##STR36## wherein R 2 and R 4 are independently hydrogen, alkyl, aryl or an electron withdrawing group having a positive Hammett sigma value, R 3 is R 1 , hydrogen, alkyl, aryl or an electron withdrawing group having a positive Hammett sigma value, or R 3 and R 4 , taken together, represent the atoms necessary to complete a fused carbocyclic ring, R 5 is alkylene of 1 or 2 carbon atoms, R 6 is methyl, Q is carbonyl or thiocarbonyl, FRAG is a fluorescent moiety derived from a compound selected from the group consisting of ##STR37## wherein R is hydroxy or mercapto or salts thereof, and m is 0 or 1, said fluorescent moiety having an oxy or thio linkage, derived from said R group by removal of a hydrogen atom, through which said moiety is attached to said CAR-- nucleus, provided that when R 1 is replaced with H, CAR--H has an E 1/2 of at least about +100 mV when measured in water.

Claim 19

The element of claim 14 further comprising a carbon source nutrient for living organisms.

Claim 22

The method of claim 16 wherein said reducible compound has the structure ##STR39## wherein R 2 and R 4 are independently hydrogen, alkyl, aryl or an electron withdrawing group having a positive Hammett sigma value, R 3 is R 1 , hydrogen, alkyl, aryl or an electron withdrawing group having a positive Hammett sigma value, or R 3 and R 4 , taken together, represent the atoms necessary to complete a fused carbocyclic ring, R 5 is alkylene of 1 or 2 carbon atoms, R 6 is methyl, Q is carbonyl or thiocarbonyl, FRAG is said fluorescent moiety attached to Q through said oxy or thio linkage, and m is 0 or 1.

Claim 23

The method of claim 16 for the determination of a living organism in the presence of an electron transfer agent.

Claim 24

The method of claim 16 wherein when said compound is reduced at about pH 7, at least about 50% of said fluorescent moiety is released within about 30 minutes.

Claim 25

The method of claim 16 wherein said fluorescent moiety is derived from ##STR40## wherein R is hydroxy or amino or salts thereof.

Claim 28

A compound of the structure ##STR42## wherein R is hydroxy, mercapto or amino or salts thereof.

Claim 29

A compound of the structure ##STR42## wherein R is hydroxy, mercapto or amino or salts thereof.