A simple solution for this potential problem is to collect the fractions of the peak, determine the OD260 in the standard way, and add that datum point to the graph (in this case, the graph may not be linear). Finally, although intuitive, it is important to point out that the usually long and thick waste eluent line connecting the UV detector flow cell to waste container needs to be removed when collecting fractions of eluent to generate the OD260-HPLC peak area correlation graph. If this were not followed, the fractions collected may not be the fractions corresponding to the intended HPLC peak.