For a biochemistry lab using absorbance, enzyme, etc. Absorbance was found for varying concentrations of p-nitrophenol at pH 8.0 and pH 14.0
I just need someone to guide me through the process of finding the "absorptivity index". What is the formula?
JUD lhe reactions by adding 1.0 mL of 2.5 M NaoH at 30 second intervals. (Don't forget to add 1 mL of 2.5 M NaoH to the blank.) Read A400 for each tube using the blank to zero the spectrophotometer. Write-Up For Experiment #6 1. Plot the absorbance of p-nitrophenol against concentration (umole/5 mL) at pH 8.0 and pH 14 (both on the same piece of graph paper). Calculate the extinction coefficients" from the slopes of the lines. The "extinction coefficients" you obtain here are in terms of umoles and a volume of 5 mL and thus are not molar extinction coefficients. The values obtained in each case could also be called an absorptivity index (E400.pH8 and E400 pH14.) 12. Plot the A400 versus time for the range finding assays. Obtain an absorbance change rate for each enzyme dilution which you measured (in terms of AA 400/min) 3. Plot A400 versus time for the continuous assay for the study on the effect of concentration on the rate of reaction Determine an absorbance change rate for each enzyme concentration (i.e.. AA400/min). 4. ine an absorbance change rate for each enzyme concentration for the
