And this absorption rate of TMB by uv vis depends on what factors
And whether the oxidizing agent of TMB is also effective on the amount of absorption at 650 wavelength
The absorbance of a solution at any given wavelength is directly proportional to concentration of the absorbing substance, as long as the concentration is not too high for other effects to come into play. The constant of proportionality is called the extinction coefficient. The absorbance is also directly proportional to the distance light travels through the solution. Overall, the relationship is called Beer's Law or the Beer-Lambert Law.
If there is more than one substance in the solution absorbing light at the given wavelength, the overall absorbance will be the sum of the absorbances of the individual components, as long as there is no interaction between the substances. If there is a chemical reaction between them, the situation is more dynamic, and the absorbance will change over time as the reaction occurs.
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