Aminde Detection Method and Materials

Abstract

Compounds linked to a solid support through a divalent linker moiety are disclosed. In particular, compounds such as 1-hydroxybenzotriazole-6-carboxylic acid are directly linked to the support under mild conditions (i.e., in aqueous or organic solvents at neutral pH and at room temperature). The polymer bound 1-hydroxybenzotriazole-6-carboxylic acid can be used for the derivatization of amines as well as for single step amino group modification of proteins, peptides, and amines via acylation or sulfonylation reactions. A flow through device and method for the single step amino group modifications of proteins, peptides, and amines is disclosed. Also disclosed is a flow through device for the detection of amines in a sample. Additionally, a device and method for the detection of amines in a sample using 1-hydroxybenzotriazole-6-carboxylic acid are disclosed. In a preferred embodiment, the device is used to detect the presence of amines in a spoiled meat product. Diagnostic kits for detecting the presence of amines are also disclosed.

1 - 78 . (canceled) 
   
   
       79 . An indicator for detecting the presence of an amine in a sample comprising an indicator releaseably bound to a solid or membrane support;
 wherein in the presence of an amine, said indicator is released from said support, thereby causing a detectable color change;   and wherein said color change is proportional to the amount of amine present in said sample.   
   
   
       80 . The indicator of  claim 79 , wherein said indicator is releaseably bound to a compound linked to a solid or membrane support, wherein said compound linked to said support is represented by the following formula: 
     
       
         
         
             
             
         
       
       wherein X represents a divalent linker moiety; 
       R represents said solid or membrane support; 
       and R′ represents a mono- or polyaromatic ring, which may include one or more heteroatoms. 
     
   
   
       81 . The indicator of  claim 80 , wherein R′ is selected from the group consisting of 1-hydroxybenzotriazole-6-carboxylic acid, 2,3,5,6-tetrafluoro-4-hydroxybenzoic acid, 4-hydroxy-3-nitrobenzoic acid, 6-hydroxynicotinic acid, and 2-hydroxy-1,3-dioxoisoindole-5-carboxylic acid, and derivatives thereof. 
   
   
       82 . The indicator of  claim 79 , wherein said support is selected from the group consisting of polystyrene, polystyrene/polyethylene glycol graft copolymers, silica gels, glass beads, controlled pore glass, agarose, sepharose, cellulose, chitosan, polyacrylonitrile, polyurethane, polypropylene, polyvinyl alcohol, polymethacrylate, polyacrylamide, polysulfone and modified derivatives thereof. 
   
   
       83 . The indicator of  claim 80 , wherein said linker moiety is selected from the group consisting of N—R″ and O; wherein R″ is H or an alkyl group. 
   
   
       84 . The indicator of  claim 80 , wherein said compound linked to said support is 1-hydroxybenzotriazole-6-carboxylic acid and is represented by the following formula: 
     
       
         
         
             
             
         
       
     
     wherein R and X are as previously described. 
   
   
       85 . The indicator of  claim 79 , wherein said releasable indicator is a dye. 
   
   
       86 . The indicator of  claim 83 , wherein said dye is FD&C Red 3. 
   
   
       87 . The indicator of  claim 85 , wherein said releasable indicator is an azo dye. 
   
   
       88 . The indicator of  claim 87 , wherein said azo dye is selected from the group consisting of DABCYL and DABSYL. 
   
   
       89 . A compound linked to a solid or membrane support, wherein said compound linked to said support is represented by the following formula: 
     
       
         
         
             
             
         
       
       
         
         
             
             
         
       
       R represents said support. 
     
   
   
       90 . The compound of  claim 89 , wherein R′ is selected from the group consisting of 1-hydroxybenzotriazole-6-carboxylic acid, 2,3,5,6-tetrafluoro-4-hydroxybenzoic acid, 4-hydroxy-3-nitrobenzoic acid, 6-hydroxynicotinic acid and 2-hydroxy-1,3-dioxoisoindole-5-carboxylic acid, and derivatives thereof. 
   
   
       91 . The compound of  claim 89 , wherein said compound linked to said support is 1-hydroxybenzotriazole-6-carboxylic acid and is represented by the following formula: 
     
       
         
         
             
             
         
       
       wherein R and X are as defined above. 
     
   
   
       92 . The compound of  claim 89 , wherein said support is selected from the group consisting of polystyrene, polystyrene/polyethylene glycol graft copolymers, silica gels, glass beads, controlled pore glass, agarose, sepharose, cellulose, chitosan, polyacrylonitrile, polyurethane, polypropylene, polyvinyl alcohol, polymethacrylate, polyacrylamide, polysulfone and modified derivatives thereof. 
   
   
       93 . The compound of  claim 89 , wherein said linker moiety is selected from the group consisting of N—R″ and O; wherein R″ is H or an alkyl group. 
   
   
       94 . The compound of  claim 89 , wherein said compound bears a detectable agent selected from the group consisting of a chromophoric agent, a fluorogenic agent, a radioactive agent and an electrochemical agent. 
   
   
       95 . The compound of  claim 94 , wherein said detecable agent is selected from the group consisting of FD&C Red 3, DABCYL, DABSYL, pyrene, fluorescein, lucifer yellow, BODIPY, rhodamine, DANSYL, EDANS, .sup.3H, .sup.14C, .sup.125I, .sup.35S, .sup.32P. 
   
   
       96 . The compound of  claim 94  wherein the electrochemical agent contains a thiol or catechol moiety. 
   
   
       97 . The compound of  claim 93 , wherein said amide or ester linkage enhances the chemical reactivity of said compound.