When I have modified gradient program in HPLC analysis, same sample shows different area response at different RT.
Hello, the by changing gradient flow you changing mobile phase composition which definitely will act on RT and shape of picks.
The peak area is not always a constant value for a measured sample of a certain concentration. The peak area is fairly constant for a given system: sample-substance-moving phase-non-moving phase, taking into account phase ratios and flow rate. If you change a component of the system, the peak data may change. You can compare the peak areas of different samples or different concentrations of the same sample under the same experimental conditions. If you change the experimental mode, you will have to start over again, obtaining the raw data for each sample and each concentration again
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