The high-performance liquid chromatography (HPLC) is one
of the kinds of chromatography used to separate the molecules using the mobile
phase and a stationary phase. Each analyte has a specific structure, pKa,
polarity, solubility, and according to this they have a different affinity
towards the stationary phase and cause different retention times.
Theoretically, when we inject the same sample of analyte
and a change flow rate, the peak area for this analysis has to be unchanged,
with the system parameters keeping everything the same. But actually, in high-pressure liquid chromatography,
the peak area increases as the flow rate decreases (vice-versa for increase
flow rate), while the same concentration of the sample injected and passes
through the detector. This is because the width of the peak will increase as
the flow rate decreases and the area of the peak will increase as a result.
This is the major effect of flow rate on peak area in HPLC.
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