I am trying to get a calibration curve of polyphenol standard, but i am getting variation in the peaks run at different concentrations, does this matter? How can i minimize this error?
If it is a new column or if the column is also used for a different method, try to condition it by running the mobile phase for a couple of hours. This might help....
Is the variation in retention time within experiment run within day or in different day?? are you running isocratic?? What is the composition of mobile phase (is it contain buffer?) Have you adjusted the temperature of experiment. If everything is fine, just double the run time of experiment with retention time of last compound eluted in chromatogram and check it. Hope it can also solve your problem. Keep sufficient equilibration of column with mobile phase, detector and oven. Good luck.
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