Use of substituted quinone electron transfer agents in analytical determinations

Claim 2

A composition for the determination of an analyte in a liquid, said composition comprising: (a) an at least partially substituted benzo- or naphthoquinone electron transfer agent (ETA) which is capable of being reduced by said analyte and has an E 1/2 of from about -320 to about +400 mV as measured in an aqueous buffer solution at pH 7, (b) a reducible compound which provides a detectable species when reduced by said reduced ETA, and (c) a buffer which maintains the pH of said composition at pH 9 or less, said reducible compound being selected from the group consisting of tetrazolium salts dichloroindophenol dyes, and compounds of the structure CAR--R 1 ) n wherein CAR-- is an aromatic or quinone nucleus, R 1 is a moiety which comprises a shiftable detectable species, and n is 1 or 2, provided that said reducible compound is capable of being reduced at a pH of 9 or less to release said shiftable detectable species, and further provided that when R 1 is replaced with H, CAR--H) n has an E1/2 of either at least about +100 mV when measured in water, or of at least about -650 mV when measured in acetonitrile.

Claim 3

A composition for the determination of an analyte in a liquid, said composition comprising: (a) an at least partially substituted benzo- or naphthoquinone electron transfer agent (ETA) which is capable of being reduced by said analyte and has an E 178 of from about -320 mV to about +400 mV as measured in buffer at pH 7, and (b) a reducible compound represented by the structure CAR--R 1 ) n wherein CAR-- is an aromatic or quinone nucleus, R 1 is a moiety which comprises a shiftable detectable species, and n is 1 or 2, provided that said reducible compound is capable of being reduced at a pH of 9 or less to release said shiftable detectable species, and further provided that when R 1 is replaced with H, CAR--H) n has an E 1/2 of either at least about +100 mV when measured in water, or of at least about -650 mV when measured in acetonitrile.

Claim 4

A composition for the determination of an analyte in a liquid, said composition comprising: (a) an at least partially substituted benzo- or naphthoquinone electron transfer agent (ETA) which is capable of being reduced by said analyte and has an E 178 of from about -320 mV to about +400 mV as measured in buffer at pH 7, and (b) a reducible compound represented by the structure CAR--R 1 ) n wherein CAR-- is an aromatic or quinone nucleus, R 1 is a moiety which comprises a shiftable detectable species, and n is 1 or 2, provided that said reducible compound is capable of being reduced at a pH of 9 or less to release said shiftable detectable species, and further provided that when R 1 is replaced with H, CAR--H) n has an E 1/2 of either at least about +100 mV when measured in water, or of at least about -650 mV when measured in acetonitrile.

Claim 5

The composition of claim 2 wherein said reducible compound is a reducible intramolecular nucleophilic displacement (RIND) compound having the structure CAR--R 1 wherein CAR-- is ##STR24## R 2 and R 4 are independently hydrogen, alkyl, aryl or an electron withdrawing group, R 3 is R 1 , hydrogen, alkyl, aryl or an electron withdrawing group, 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 alkyl, cycloalkyl or aryl, Q is carbonyl or thiocarbonyl, FRAG is a shiftable detectable species which provides a detectable species when released from said reducible compound, and m is 0 or 1, provided when R 1 is replaced with H, CAR--H has an E 1/2 of either at least about +100 mV when measured in water, or at least about -650 mV when measured in acetonitrile.

Claim 6

The composition of claim 2 wherein said ETA has the structure ##STR25## wherein R 7 and R 8 are independently hydrogen, alkyl, alkenyl, alkoxy, hydroxyalkyl, hydroxyalkoxy, alkoxyalkyl, alkoxyalkoxy, acetoxyalkyl, acetoxyalkoxy, aryl, alkaryl, a heterocycle or an heteroalkyl, and R 9 and R 10 are independently selected from the substituents defined as R 7 or R 8 , or taken together supply the atoms to complete a 4- to 8-membered fused carbocyclic or heterocyclic ring, provided at least one of R 7 , R 8 , R 9 and R 10 is not hydrogen.

Claim 7

The composition of claim 4 wherein R 7 and R 8 are independently hydrogen, alkyl, alkoxy or hydroxyalkyl, and R 9 and R 10 are independently selected from the substituents defined as R 7 or R 8 , or taken together, supply the atoms to complete a 6- to 8-membered fused carbocyclic ring, and said ETA has an E 1/2 of from about -185 mV to about +400 mV as measured in buffer at pH 7.

Claim 8

The composition of claim 2 wherein said ETA is 2,3,5-trimethyl-1,4-benzoquinone, 2,3-dimethyl-5-hydroxymethyl-1,4-benzoquinone or 2,3-dimethoxy-5-methyl-1,4-benzoquinone.

Claim 9

The composition of claim 3 wherein m is 1, at least two of R 2 , R 3 and R 4 are independently electron withdrawing groups, or R 3 and R 4 , taken together, represent the atoms necessary to complete a fused 5- to 7-membered carbocyclic ring, R 5 is methylene, R 6 is alkyl of 1 to 3 carbon atoms or phenyl, Q is carbonyl, and FRAG is a chromogen or fluorogen.

Claim 10

A dry analytical element for the determination of an analyte in a liquid, said element comprising an absorbent carrier material, an at least partially substituted benzo- or naphthoquinone electron transfer agent (ETA) which is capable of being reduced by said analyte and has an E 1/2 of from about -320 mV to about +400 mV as measured in buffer at pH 7, and a reducible compound which provides a detectable species when reduced by said reduced ETA.

Claim 11

A dry analytical element for the determination of an analyte in a liquid, said element comprising an absorbent carrier material, an at least partially substituted benzo- or naphthoquinone electron transfer agent (ETA) which is capable of being reduced by said analyte and has an E 1/2 of from about -320 mV to about +400 mV as measured in buffer at pH 7, and a reducible compound which provides a detectable species when reduced by said reduced ETA.

Claim 12

The element of claim 8 wherein said ETA has the structure ##STR26## wherein R 7 and R 8 are independently hydrogen, alkyl, alkenyl, alkoxy, hydroxyalkyl, hydroxyalkoxy, alkoxyalkyl, alkoxyalkoy, acetoxyalkyl, acetoxyalkoxy, aryl, alkaryl, a heterocycle or an heteroalkyl, and R 9 and R 10 are independently selected from the substituents defined as R 7 or R 8 , or taken together supply the atoms to complete a 4- to 8-membered fused carbocyclic or heterocyclic ring, provided at least one of R 7 , R 8 , R 9 and R 10 is not hydrogen, and said reducible compound is a reducible intramolecular nucleophilic displacement (RIND) compound having the structure CAR--R 1 wherein CAR-- is ##STR27## R 2 and R 4 are independently hydrogen, alkyl, aryl or an electron withdrawing group, R 3 is R 1 , hydrogen, alkyl, aryl or an electron withdrawing group, 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 alkyl, cycloalkyl or aryl, Q is carbonyl or thiocarbonyl, FRAG is a shiftable detectable species which provides a detectable species when released from said reducible compound, and m is 0 or 1, provided when R 1 is replaced with H, CAR--H has an E 1/2 of either at least about +100 mV when measured in water, or at least about -650 mV when measured in acetonitrile.

Claim 13

The element of claim 9 wherein at least two of R 2 , R 3 and R 4 are independently electron withdrawing groups, or R 3 and R 4 , taken together, represent the atoms necessary to complete a fused 5- to 7-membered carboxylic ring, R 5 is methylene, R 6 is alkyl of 1 to 3 carbon atoms or phenyl, Q is carbonyl, FRAG is a chromogen or fluorogen, R 7 and R 8 are independently hydrogen, alkyl, alkoxy or hydroxyalkyl, and R 9 and R 10 are independently selected from the substituents defined as R 7 or R 8 , or taken together, supply the atoms to complete a 6- to 8-membered fused carbocyclic ring, and said ETA has an E 1/2 of from about -185 mV to about +400 mV measured in buffer at pH 7.

Claim 14

The element of claim 8 comprising an interactive composition for said analyte.

Claim 15

The element of claim 8 comprising a carbon source nutrient for living organisms.

Claim 18

The method of claim 13 to determine a non-living analyte in the presence of an interactive composition for said analyte.

Claim 19

The method of claim 13 to determine bacterial cells.

Claim 20

The method of claim 13 wherein said reducible compound is represented by the structure CAR--R 1 ) n wherein CAR-- is an aromatic or quinone nucleus, R 1 is a moiety which comprises a shiftable detectable species, and n is 1 or 2, provided that said reducible compound is capable of being reduced at a pH of 9 or less to release said shiftable detectable species, and further provided that when R 1 is replaced with H, CAR--H) n has an E 1/2 of either at least about +100 mV when measured in water, or of at least about -650 mV when measured in acetonitrile.

Claim 21

The method of claim 16 wherein said reducible compound is a reducible intramolecular nucleophilic displacement (RIND) compound having the structure CAR--R 1 wherein CAR is ##STR28## R 2 and R 4 are independently hydrogen, alkyl, aryl or an electron withdrawing group, R 3 is R 1 , hydrogen, alkyl, aryl or an electron withdrawing group, 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 alkyl, cycloalkyl or aryl, Q is carbonyl or thiocarbonyl, FRAG is a shiftable detectable species which provides a detectable species when released from said reducible compound, and m is 0 or 1, provided when R 1 is replaced with H, CAR--H has an E 1/2 of either at least about +100 mV when measured in water, or at least about -650 mV when measured in acetonitrile, and said ETA has the structure ##STR29## wherein R 7 and R 8 are independently hydrogen, alkyl, alkenyl, alkoxy, hyroxyalkyl, hydroxyalkoxy, alkoxyalkyl, alkoxyalkoxy, acetoxyalkyl, acetoxyalkoxy, aryl, alkaryl, a heterocycle or an heteroalkyl, and R 9 and R 10 are independently selected from the substituents defined as R 7 or R 8 , or taken together supply the atoms to complete a 4- to 8-membered fused carbocyclic or heterocyclic ring, provided at least one of R 7 , R 8 , R 9 and R 10 is not hydrogen.

Claim 22

The method of claim 17 wherein R 7 and R 8 are independently hydrogen, alkyl, alkoxy or hydroxyalkyl, and R 9 and R 10 are independently selected from the substituents defined as R 7 or R 8 , or taken together, supply the atoms to complete a 6- to 8-membered fused carbocyclic ring, provided said ETA has an E 1/2 of from about -185 mV to about +400 mV as measured in an aqueous buffer solution at pH 7.

Claim 23

The method of claim 13 wherein step B is carried out within 60 minutes of step A.

Claim 24

A composition for the determination of an analyte in a liquid, said composition comprising: (a) an at least a partially substituted naphthoquinone electron transfer agent (ETA) which is capable of being reduced by said analyte and has an E 1/2 of from about -320 to about +400 mV as measured in an aqueous buffer solution at pH 7, said naphthoquinone being represented by the structure: ##STR30## wherein SUB represents one or more electron withdrawing substituents, electron releasing substituents, or substituents which provide an aromatic or nonaromatic carbocyclic or heterocyclic ring fused with the naphthoquinone nucleus to provide said ETA with the desired E 1/2 value, (b) a reducible compound which provides a detectable species when reduced by said reduced ETA, and (c) a buffer which maintains the pH of said composition at pH 9 or less.

Claim 25

A composition for the determination of an analyte in a liquid, said composition comprising: (a) an at least a partially substituted naphthoquinone electron transfer agent (ETA) which is capable of being reduced by said analyte and has an E 1/2 of from about -320 to about +400 mV as measured in an aqueous buffer solution at pH 7, said naphthoquinone being represented by the structure: ##STR30## wherein SUB represents one or more electron withdrawing substituents, electron releasing substituents, or substituents which provide an aromatic or nonaromatic carbocyclic or heterocyclic ring fused with the naphthoquinone nucleus to provide said ETA with the desired E 1/2 value, (b) a reducible compound which provides a detectable species when reduced by said reduced ETA, and (c) a buffer which maintains the pH of said composition at pH 9 or less.