Question

1. Two fluorescent compounds A and B are present in a mixture. Using a spectrofluorimeter similar to the one used in this experiment, how would you determine the quantity of A in the mixture if-

A absorbs radiation in a region where B does not.

Both A and B absorb in the same region but A emits fluorescence at different wavelengths than B.

Answer 1

Case 1: When A absorbs radiation in region where B does not

In this case, we take/ measure the samples in the same region at the given wavelength. We then identify which peak will correspond to Compund A depending on its struture. Measuring the intensity, then will be proportional to the quantity of the compund A if done for low concentrations.

Case 2: Both A and B absorb in same region but at different wavelengths.

This is easy as already, we can identity at different wavelengths , which peak will correspond to what in the given region. Hence, the known intensity of peak will decipher the quantity of A.

Have a nice day! :)