Method for assaying the antioxidant capacity of a sample

Abstract

A kit for assaying the antioxidant capacity of a sample, the kit including an extraction solution including a solubility enhancing compound to be added to a sample for extracting antioxidants present in the sample; and a fluorescent probe to be added to the extract.

1 . A kit for assaying the antioxidant capacity of a sample, the kit including: 
 an extraction solution including a solubility enhancing compound to be added to a sample for extracting antioxidants present in the sample; and    a fluorescent probe to be added to the extract.    
     
     
         2 . The kit of  claim 1  in which the extraction solution includes a high polarity solvent and a low polarity solvent.  
     
     
         3 . The kit of  claim 2  in which the high polarity solvent is water.  
     
     
         4 . The kit of  claim 2  in which the low polarity solvent is selected from the group consisting of acetone, butanone, methanol, acetonitrile, methylene chloride, 1-2-dichloroethane.  
     
     
         5 . The kit of  claim 1  in which the solubility enhancing compound is cyclodextrin and derivatives thereof.  
     
     
         6 . The kit of  claim 2  in which the amount of the high polarity solvent is equal to or approximately equal to the amount of low polarity solvent.  
     
     
         7 . The kit of  claim 1  in which the solubility enhancing compound is 1% to 40% of the solution.  
     
     
         8 . The kit of  claim 1  in which the probe is a non-protein probe.  
     
     
         9 . The kit of  claim 8  in which the non-protein probe is a hydrogen atom donor probe.  
     
     
         10 . The kit of  claim 9  in which the hydrogen atom donor probe is fluorescein.  
     
     
         11 . The kit of  claim 1  further including: 
 a plurality of standards each having a known antioxidant capacity so that the fluorescence intensity decay of the probe in the presence of each standard over time can be detected.    
     
     
         12 . The kit of  claim 11  in which there are four standards.  
     
     
         13 . The kit of  claim 11  in which the concentration of the standards ranges from 10 mM to 100 mM.  
     
     
         14 . The kit of  claim 11  in which each standard is Trolox.  
     
     
         15 . The kit of  claim 1  further including adding a free radical generator precursor to be added to the probe/extract mixture.  
     
     
         16 . The kit of  claim 15  in which the precursor is AAPH.  
     
     
         17 . The kit of  claim 15  in which the concentration of the precursor is above 4 mM.  
     
     
         18 . The kit of  claim 17  in which the concentration of the precursor is 12 mM.  
     
     
         19 . A kit for assaying the antioxidant capacity of a sample, the kit comprising: 
 an extraction solution to be added to a sample for extracting antioxidants present in the solution; and    a non-protein fluorescent probe to be added to the extract.    
     
     
         20 . The kit of  claim 19  in which the extraction solution includes a high polarity solvent and a low polarity solvent.  
     
     
         21 . The kit of  claim 20  in which the high polarity solvent is water.  
     
     
         22 . The kit of  claim 20  in which the low polarity solvent is selected from the group consisting of acetone, butanone, methanol, acetonitrile, methylene chloride, 1,2-dichloroethane, and ethanol.  
     
     
         23 . The kit of  claim 20  in which the amount of the high polarity solvent is equal to or approximately equal to the amount of low polarity solvent.  
     
     
         24 . The kit of  claim 19  further including a solubility enhancing compound to be added to the extraction solution.  
     
     
         25 . The kit of  claim 24  in which the solubility enhancing compound is cyclodextrine and derivatives thereof.  
     
     
         26 . The kit of  claim 24  in which the solubility enhancing compound is 1% to 40% of the extraction solution.  
     
     
         27 . The kit of  claim 19  in which the non-protein probe is a hydrogen atom donor probe.  
     
     
         28 . The kit of  claim 27  in which the hydrogen atom donor probe is fluorescein.  
     
     
         29 . The kit of  claim 19  further including a plurality of standards each having a known antioxidant capacity so that the fluorescence intensity decay of the probe in the presence of each standard over time can be detected.  
     
     
         30 . The kit of  claim 29  in which there are four standards.  
     
     
         31 . The kit of  claim 30  in which the concentration of the standards ranges from 10 mM moles to 100 mM moles.  
     
     
         32 . The kit of  claim 29  in which each standard is Trolox.  
     
     
         33 . The kit of  claim 19  further including a free radical generator precursor to be added to the probe/extract mixture.  
     
     
         34 . The kit of  claim 33  in which the precursor is AAPH.  
     
     
         35 . The kit of  claim 33  in which the concentration of the precursor is above 4 mM.  
     
     
         36 . The kit of  claim 35  in which the concentration of the precursor is 12.8.  
     
     
         37 . A kit for assaying the antioxidant capacity of a sample, the kit including: 
 an extraction solution to be added to a sample for extracting antioxidants present in the sample; and    a fluorescent probe to be added to the extract.    
     
     
         38 . The kit of  claim 37  in which the extraction solution includes a high polarity solvent and a low polarity solvent.  
     
     
         39 . The kit of  claim 38  in which the high polarity solvent is water.  
     
     
         40 . The kit of  claim 38  in which the low polarity solvent is selected from the group consisting of acetone, butanone, methanol, acetonitrile, methylene chloride, 1-2-dichloroethane.  
     
     
         41 . The kit of  claim 38  in which the amount of the high polarity solvent is equal to or approximately equal to the amount of low polarity solvent.  
     
     
         42 . The kit of  claim 37  further including: 
 a plurality of standards each having a known antioxidant capacity so that the fluorescence intensity decay of the probe in the presence of each standard over time can be detected.    
     
     
         43 . The kit of  claim 42  in which there are four standards.  
     
     
         44 . The kit of  claim 42  in which the concentration of the standards ranges from 10 mM to 100 mM.  
     
     
         45 . The kit of  claim 42  in which each standard is Trolox.  
     
     
         46 . The kit of  claim 37  further including adding a free radical generator precursor to be added to the probe/extract mixture.  
     
     
         47 . The kit of  claim 46  in which the precursor is AAPH.  
     
     
         48 . The kit of  claim 46  in which the concentration of the precursor is above 4 mM.  
     
     
         49 . The kit of  claim 48  in which the concentration of the precursor is 12 mM.