Wednesday, October 2, 2019

Effect of Flow Rate on Peak Area in HPLC

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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