Methods and kits for detecting hemoglobin in test samples

Abstract

The present invention relates to methods of detecting hemoglobin in a test sample. These methods can be used to diagnose a subject suffering from a genetic disorder relating to hemoglobin metabolism, to determine the eligibility of a subject to be a blood donor, to determine the age of a stored blood sample or to identify a hemolyzed plasma sample. The present invention also relates to kits for use in the above described methods.

1 .- 43 . (canceled) 
     
     
         44 : A kit for detecting hemoglobin in a test sample, the kit comprising:
 a. at least one basic solution;   b. at least one indicator solution, wherein the indicator solution comprises at least one acridinium compound; and   c. instructions for detecting hemoglobin in a test sample.   
     
     
         45 : The kit of  claim 44 , wherein the basic solution is a solution having a pH of at least about 10. 
     
     
         46 : The kit of  claim 44 , wherein the acridinium compound is an acridinium-9-carboxamide having a structure according to formula I: 
       
         
           
           
               
               
           
         
         wherein R 1  and R 2  are each independently selected from the group consisting of: alkyl, alkenyl, alkynyl, aryl or aralkyl, sulfoalkyl, carboxyalkyl and oxoalkyl, and 
         wherein R 3  through R 15  are each independently selected from the group consisting of: hydrogen, alkyl, alkenyl, alkynyl, aryl or aralkyl, amino, amido, acyl, alkoxyl, hydroxyl, carboxyl, halogen, halide, nitro, cyano, sulfo, sulfoalkyl, carboxyalkyl and oxoalkyl; and optionally, if present, X ⊖  is an anion. 
       
     
     
         47 : The kit of  claim 44 , wherein the acridinium compound is an acridinium-9-carboxylate aryl ester having a structure according to formula II: 
       
         
           
           
               
               
           
         
         wherein R 1  is an alkyl, alkenyl, alkynyl, aryl or aralkyl, sulfoalkyl, carboxyalkyl and oxoalkyl; and 
         wherein R 3  through R 15  are each independently selected from the group consisting of: hydrogen, alkyl, alkenyl, alkynyl, aryl or aralkyl, amino, amido, acyl, alkoxyl, hydroxyl, carboxyl, halogen, halide, nitro, cyano, sulfo, sulfoalkyl, carboxyalkyl and oxoalkyl; and optionally, if present, X ⊖  is an anion. 
       
     
     
         48 : The kit of  claim 44 , wherein the indicator solution further comprises at least one surfactant. 
     
     
         49 - 50 . (canceled) 
     
     
         51 : A kit for diagnosing a subject suffering from a genetic disorder relating to hemoglobin metabolism, the kit comprising:
 a. at least one basic solution;   b. at least one indicator solution, wherein the indicator solution comprises at least one acridinium compound; and   c. instructions for diagnosing a subject suffering from a genetic disorder relating to hemoglobin metabolism.   
     
     
         52 : The kit of  claim 51 , wherein the basic solution is a solution having a pH of at least about 10. 
     
     
         53 : The kit of  claim 51 , wherein the acridinium compound is an acridinium-9-carboxamide having a structure according to formula I: 
       
         
           
           
               
               
           
         
         wherein R 1  and R 2  are each independently selected from the group consisting of alkyl, alkenyl, alkynyl, aryl or aralkyl, sulfoalkyl, carboxyalkyl and oxoalkyl, and 
         wherein R 3  through R 15  are each independently selected from the group consisting of: hydrogen, alkyl, alkenyl, alkynyl, aryl or aralkyl, amino, amido, acyl, alkoxyl, hydroxyl, carboxyl, halogen, halide, nitro, cyano, sulfo, sulfoalkyl, carboxyalkyl and oxoalkyl; and optionally, if present, X ⊖  is an anion. 
       
     
     
         54 : The kit of  claim 51 , wherein the acridinium compound is an acridinium-9-carboxylate aryl ester having a structure according to formula II: 
       
         
           
           
               
               
           
         
         wherein R 1  is an alkyl, alkenyl, alkynyl, aryl or aralkyl, sulfoalkyl, carboxyalkyl and oxoalkyl; and 
         wherein R 3  through R 15  are each independently selected from the group consisting of: hydrogen, alkyl, alkenyl, alkynyl, aryl or aralkyl, amino, amido, acyl, alkoxyl, hydroxyl, carboxyl, halogen, halide, nitro, cyano, sulfo, sulfoalkyl, carboxyalkyl and oxoalkyl; and optionally, if present, X ⊖  is an anion. 
       
     
     
         55 : The kit of  claim 51 , wherein the indicator solution further comprises at least one surfactant. 
     
     
         56 : A kit for determining the eligibility of a subject to be a blood donor, the kit comprising:
 a. at least one basic solution;   b. at least one indicator solution, wherein the indicator solution comprises at least one acridinium compound; and   c. instructions for determining the eligibility of a subject to be a blood donor.   
     
     
         57 : The kit of  claim 56 , wherein the basic solution is a solution having a pH of at least about 10. 
     
     
         58 : The kit of  claim 56 , wherein the acridinium compound is an acridinium-9-carboxamide having a structure according to formula I: 
       
         
           
           
               
               
           
         
         wherein R 1  and R 2  are each independently selected from the group consisting of: alkyl, alkenyl, alkynyl, aryl or aralkyl, sulfoalkyl, carboxyalkyl and oxoalkyl, and 
         wherein R 3  through R 15  are each independently selected from the group consisting of: hydrogen, alkyl, alkenyl, alkynyl, aryl or aralkyl, amino, amido, acyl, alkoxyl, hydroxyl, carboxyl, halogen, halide, nitro, cyano, sulfo, sulfoalkyl, carboxyalkyl and oxoalkyl; and optionally, if present, X ⊖  is an anion. 
       
     
     
         59 : The kit of  claim 56 , wherein the acridinium compound is an acridinium-9-carboxylate aryl ester having a structure according to formula II: 
       
         
           
           
               
               
           
         
         wherein R 1  is an alkyl, alkenyl, alkynyl, aryl or aralkyl, sulfoalkyl, carboxyalkyl and oxoalkyl; and 
         wherein R 3  through R 15  are each independently selected from the group consisting of: hydrogen, alkyl, alkenyl, alkynyl, aryl or aralkyl, amino, amido, acyl, alkoxyl, hydroxyl, carboxyl, halogen, halide, nitro, cyano, sulfo, sulfoalkyl, carboxyalkyl and oxoalkyl; and optionally, if present, X ⊖  is an anion. 
       
     
     
         60 : The kit of  claim 56 , wherein the indicator solution further comprises at least one surfactant. 
     
     
         61 : A kit for determining the age of a stored blood sample, the kit comprising:
 a. at least one basic solution;   b. at least one indicator solution, wherein the indicator solution comprises at least one acridinium compound; and   c. instructions for determining the age of a blood sample.   
     
     
         62 : The kit of  claim 61 , wherein the basic solution is a solution having a pH of at least about 10. 
     
     
         63 : The kit of  claim 61 , wherein the acridinium compound is an acridinium-9-carboxamide having a structure according to formula I: 
       
         
           
           
               
               
           
         
         wherein R 1  and R 2  are each independently selected from the group consisting of: alkyl, alkenyl, alkynyl, aryl or aralkyl, sulfoalkyl, carboxyalkyl and oxoalkyl, and 
         wherein R 3  through R 15  are each independently selected from the group consisting of: hydrogen, alkyl, alkenyl, alkynyl, aryl or aralkyl, amino, amido, acyl, alkoxyl, hydroxyl, carboxyl, halogen, halide, nitro, cyano, sulfo, sulfoalkyl, carboxyalkyl and oxoalkyl; and optionally, if present, X ⊖  is an anion. 
       
     
     
         64 : The kit of  claim 61 , wherein the acridinium compound is an acridinium-9-carboxylate aryl ester having a structure according to formula II: 
       
         
           
           
               
               
           
         
         wherein R 1  is an alkyl, alkenyl, alkynyl, aryl or aralkyl, sulfoalkyl, carboxyalkyl and oxoalkyl; and 
         wherein R 3  through R 15  are each independently selected from the group consisting of: hydrogen, alkyl, alkenyl, alkynyl, aryl or aralkyl, amino, amido, acyl, alkoxyl, hydroxyl, carboxyl, halogen, halide, nitro, cyano, sulfo, sulfoalkyl, carboxyalkyl and oxoalkyl; and optionally, if present, X ⊖  is an anion. 
       
     
     
         65 : The kit of  claim 61 , wherein the indicator solution further comprises at least one surfactant. 
     
     
         66 : A kit for identifying a hemolyzed serum or plasma sample, the kit comprising:
 a. at least one basic solution;   b. at least one indicator solution, wherein the indicator solution comprises at least one acridinium compound; and   c. instructions for identifying a hemolyzed serum or plasma sample.   
     
     
         67 : The kit of  claim 66 , wherein the basic solution is a solution having a pH of at least about 10. 
     
     
         68 : The kit of  claim 66 , wherein the acridinium compound is an acridinium-9-carboxamide having a structure according to formula I: 
       
         
           
           
               
               
           
         
         wherein R 1  and R 2  are each independently selected from the group consisting of: alkyl, alkenyl, alkynyl, aryl or aralkyl, sulfoalkyl, carboxyalkyl and oxoalkyl, and 
         wherein R 3  through R 15  are each independently selected from the group consisting of: hydrogen, alkyl, alkenyl, alkynyl, aryl or aralkyl, amino, amido, acyl, alkoxyl, hydroxyl, carboxyl, halogen, halide, nitro, cyano, sulfo, sulfoalkyl, carboxyalkyl and oxoalkyl; and optionally, if present, X ⊖  is an anion. 
       
     
     
         69 : The kit of  claim 66 , wherein the acridinium compound is an acridinium-9-carboxylate aryl ester having a structure according to formula II: 
       
         
           
           
               
               
           
         
         wherein R 1  is an alkyl, alkenyl, alkynyl, aryl or aralkyl, sulfoalkyl, carboxyalkyl and oxoalkyl; and 
         wherein R 3  through R 15  are each independently selected from the group consisting of: hydrogen, alkyl, alkenyl, alkynyl, aryl or aralkyl, amino, amido, acyl, alkoxyl, hydroxyl, carboxyl, halogen, halide, nitro, cyano, sulfo, sulfoalkyl, carboxyalkyl and oxoalkyl; and optionally, if present, X ⊖  is an anion. 
       
     
     
         70 : The kit of  claim 66 , wherein the indicator solution further comprises at least one surfactant.