It is the physical properties of the target molecules that determine what the most suitable HPLC column is for analytes separation in the lab. When selecting the HPLC column, the key characters you should pay attention to include hydrophobicity or hydrophilicity, intermolecular forces, intramolecular forces, and size.
In the molecular properties of the target molecules, multiple differences can be exploited by HPLC column separations. And the analyte elution profile is determined by the structure and chemistry of the HPLC column packing (stationary phase).
From the capillary to process scale, we can divide the HPLC column into different sizes. The quantity of sample loaded onto a column and the sensitivity of separation indicate the internal diameter and volume of an HPLC column. The column internal diameter can affect the separation profile. When you use gradient elution, the smaller internal diameter will increase separation and detection sensitivity. For analytical separations, we are always trying to find the balancing point between sensitivity and the sample volume loaded onto a column.
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