Unexpected Peaks for Water Sample and other Issues
Hello. I've been trying to run some samples for different concentrations of a 1-Hydroxypyrene with Methanol solution. However, the water sample that I've also been using as a reference / control of sorts has been giving undesired peaks which makes it hard to determine the appropriate data for our 1-OHP samples. The water, which I'm doing three injections of, is showing small peaks at later retention times when I would expect it to be flat the whole way through. I do have to scale a bit to see them clearly but they are noticeable even without scaling. Could this be due to A.) bad water, B.) bad solvent, or C.) bad column? Also, I was wondering if someone could answer which factors (peak width, threshold, etc.) affect integrated peak area. For the samples that are twice as concentrated as the previous, wouldn't we expect the area to be twice as large? Because that is not what we're getting. They're either the same size or slightly larger. Basically, could someone clue me in as to what the retention peaks for both water and a 1-OHP solution are supposed to look like, or is it different for each run based on things like temperature, flow rate, etc.