Q.5. When using the enzyme coupled reaction for the glucose assay, it is importa
ID: 590777 • Letter: Q
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Q.5. When using the enzyme coupled reaction for the glucose assay, it is important that the absorbance value determined at 415 nm has a direct correlation to the amount of glucose in the solution. Glucose + 02 H2ooxidass Gluconic Acid + H202 H202 ABTS(reduced) peroxidase--> ABTS,t (oxidized) (colourless) (blue) After reviewing the glucose oxidase/peroxidase-coupled reaction above discuss the following (a) How would your unknown glucose result be affected if by some chance the enzyme, catalase, was a contaminant in your unknown glucose sample? Explain your reasoning. (b) How would your unknown glucose result be affected if by some chance Vitamin C was a contaminant in your unknown glucose sample? Explain your reasoning.Explanation / Answer
a)
Catalase is an antioxidant that makes the hydrogen peroxide less toxic by converting it into water and oxygen gas. Absence of free hydrogen peroxide to oxidise the ABTS would not give the blue color.
b)
Vitamin C also serves as antioxidant by reducing the hydrogen peroxide. Therefore, no blue color of the oxidised ABTS would be produced.
Hence, in both cases, presence of glucose would be masked by antioxidant activities of the catalase and vitamin C.
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