I am analyzing a sample using size exclusion chromatography as the separation technique, and UV-VIS is used as the detector. I have some troubles regarding the calculation of the final fractions that I get from the separation. Will these two experiments give the same absorbance results?
Experiment 1: Inject 0.5 ml of sample A into the column
Experiment 2: Inject a mixture (1 ml) of 0.5 ml of sample A + 0.5 ml of solvent that does not have absorbance at the wavelength I am using.
I am aware that the absorbance is of course related to the concentration. However, in this case, both experiments have the same amount of sample A running through the column. Therefore, even though the latter is diluted, should they both give similar absorbance results or one would be 50% less? Please help to clarify.
Thank you very much in advance!
Edited by thalia12, 26 January 2017 - 07:05 PM.