Signal amplifying cobalt (III) redox reagents and methods for the determination of analytes in aqueous fluids
First Claim
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1. A method for the detection of an analyte in an aqueous sample wherein said analyte is either itself a reductant or is capable of producing a reductant comprising the steps of:
- (a) contacting said sample with reagents which effect in the presence of said analyte a sequence of reactions wherein;
(1) a water soluble cobalt(III) complex is reduced to a water soluble cobalt(II) complex by said reductant,(2) the cobalt(II) complex reacts with a water soluble dye to form a cobalt(II)-dye complex and(3) the cobalt(II)-dye complex reacts with the cobalt(III) complex to produce a cobalt(III)-dye complex and the cobalt(II) complex,(b) detecting the cobalt(III)-dye complex and relating said detected complex to said analyte.
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Abstract
Reagents and methods for the determination of analytes are disclosed, using water soluble cobalt(III) complexes and metallizable dyes to form cobalt(III) complexes of the metallizable dyes. Analytes which can be determined include enzymes, cells and biological reductants, such as NADH, NADPH and ascorbate. Amplification is provided since one equivalent of the analyte provides more than one equivalent of the detectable species.
38 Citations
29 Claims
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1. A method for the detection of an analyte in an aqueous sample wherein said analyte is either itself a reductant or is capable of producing a reductant comprising the steps of:
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(a) contacting said sample with reagents which effect in the presence of said analyte a sequence of reactions wherein; (1) a water soluble cobalt(III) complex is reduced to a water soluble cobalt(II) complex by said reductant, (2) the cobalt(II) complex reacts with a water soluble dye to form a cobalt(II)-dye complex and (3) the cobalt(II)-dye complex reacts with the cobalt(III) complex to produce a cobalt(III)-dye complex and the cobalt(II) complex, (b) detecting the cobalt(III)-dye complex and relating said detected complex to said analyte. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 23)
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14. A reagent composition for the detection of an analyte in an aqueous sample wherein said analyte is either itself a reductant or is capable of producing a reductant said composition comprising:
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(1) a water soluble cobalt(III) complex which is capable of being reduced to a water soluble cobalt(II) complex by said reductant and, (2) a water soluble dye which is capable of being metallized by said cobalt(II) complex to form a water soluble cobalt(II)-dye complex, wherein said cobalt(II)-dye complex is capable of reacting with said cobalt(III) complex to produce a cobalt(III)-dye complex and the cobalt(II) complex. - View Dependent Claims (15, 16, 17, 18, 19, 20, 21, 22, 24, 25, 26, 27)
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28. A dry element comprising an absorbent carrier material containing a composition for the detection of an analyte in an aqueous sample wherein said analyte is either itself a reductant or is capable of producing a reductant said composition comprising:
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(1) a water soluble cobalt(III) complex which is capable of being reduced to a water soluble cobalt(II) complex by said reductant and, (2) a water soluble dye which is capable of being metallized by said cobalt(II) complex to form a water soluble cobalt(II)-dye complex, wherein said cobalt(II)-dye complex is capable of reacting with said cobalt(III) complex to produce a cobalt(III)-dye complex and the cobalt(II) complex. - View Dependent Claims (29)
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Specification