Method and apparatus for determining substrate concentration and reagent for determining substrate concentration

Abstract

The present invention relates to a method, a reagent and an apparatus for determining a substrate concentration based on an amount of hydrogen peroxide generated from a substrate. In the present invention, a suppressing agent for suppressing a reaction between the hydrogen peroxide and an inhibitor is added. As the suppressing agent, an azide compound such as sodium azide or a nitrite compound such as sodium nitrite is used. In the invention, a supporting electrolyte, such as sodium chloride or potassium chloride may be further added.

1 . A method for determining a substrate concentration based on an amount of hydrogen peroxide generated from a substrate, wherein a suppressing agent for suppressing a reaction between the hydrogen peroxide and an inhibitor is allowed to coexist. 
   
   
       2 . The method for determining a substrate concentration according to  claim 1 , wherein the inhibitor is hemoglobin, and the suppressing agent is a compound which transforms the hemoglobin into methemoglobin. 
   
   
       3 . The method for determining a substrate concentration according to  claim 2 , wherein the suppressing agent is an azide compound. 
   
   
       4 . The method for determining a substrate concentration according to  claim 3 , wherein the azide compound is sodium azide. 
   
   
       5 . The method for determining a substrate concentration according to  claim 3 , wherein a concentration of the azide compound is 0.001 to 1.000 wt %. 
   
   
       6 . The method for determining a substrate concentration according to  claim 2 , wherein the suppressing agent is at least one selected from nitrites, ferricyanides, peroxides or permanganates. 
   
   
       7 . The method for determining a substrate concentration according to  claim 6 , wherein the suppressing agent is at least one selected from sodium nitrite, potassium ferricyanide, barium peroxide, or potassium permanganate. 
   
   
       8 . The method for determining a substrate concentration according to  claim 1 , wherein a supporting electrolyte is further allowed to coexist. 
   
   
       9 . The method for determining a substrate concentration according to  claim 8 , wherein the supporting electrolyte is sodium chloride or potassium chloride. 
   
   
       10 . The method for determining a substrate concentration according to  claim 8 , wherein a concentration of the supporting electrolyte is 0.01 to 2.00 wt %. 
   
   
       11 . An apparatus for determining a substrate concentration, the apparatus comprising a hydrogen peroxide electrode for determining a concentration of hydrogen peroxide generated from a substrate, wherein a suppressing agent for suppressing a reaction between the hydrogen peroxide and an inhibitor is supplied to the apparatus. 
   
   
       12 . The apparatus for determining a substrate concentration according to  claim 11 , wherein the inhibitor is hemoglobin and a compound which transforms the hemoglobin into methemoglobin is supplied as the suppressing agent. 
   
   
       13 . The apparatus for determining a substrate concentration according to  claim 12 , wherein the compound is an azide compound. 
   
   
       14 . The apparatus for determining a substrate concentration according to  claim 13 , wherein the azide compound is sodium azide. 
   
   
       15 . The apparatus for determining a substrate concentration according to  claim 12 , wherein the suppressing agent is at least one selected from nitrites, ferricyanides, peroxides or permanganates. 
   
   
       16 . The apparatus for determining a substrate concentration according to  claim 15 , wherein the suppressing agent is at least one selected from sodium nitrite, potassium ferricyanide, barium peroxide or potassium permanganate. 
   
   
       17 . The apparatus for determining a substrate concentration according to  claim 11 , wherein a supporting electrolyte is further supplied. 
   
   
       18 . A reagent for determining a substrate concentration based on an amount of hydrogen peroxide generated from a substrate, the reagent comprising a suppressing agent for suppressing a reaction between the hydrogen peroxide and an inhibitor. 
   
   
       19 . The reagent for determining a substrate concentration according to  claim 18 , wherein the inhibitor is hemoglobin, and the suppressing agent is a compound which transforms the hemoglobin into methemoglobin. 
   
   
       20 . The reagent for determining a substrate concentration according to  claim 19 , wherein the suppressing agent is an azide compound. 
   
   
       21 . The reagent for determining a substrate concentration according to  claim 20 , wherein the azide compound is sodium azide. 
   
   
       22 . The reagent for determining a substrate concentration according to  claim 19 , wherein the suppressing agent is at least one selected from nitrites, ferricyanides, peroxides, or permanganates. 
   
   
       23 . The reagent for determining a substrate concentration according to  claim 22 , wherein the nitrite is at least one selected from sodium nitrite, potassium ferricyanide, barium peroxide, or potassium permanganate. 
   
   
       24 . The reagent for determining a substrate concentration according to  claim 18 , further comprising a supporting electrolyte. 
   
   
       25 . The reagent for determining a substrate concentration according to  claim 24 , wherein the supporting electrolyte is sodium chloride or potassium chloride.