Step 3

We have established that your compound is hydrophilic and does not have a charge within pH 1.5-7.0. Below is an example of a separation of a similar compound. This example can be used a starting condition in your method development.

Hydrophilic neutral compounds can be analyzed on any Primesep column in normal mode. Since hydrophilic molecules have no or very low interaction with the stationary phase by reverse mechanism, it is practical to use the polar properties of the analytes to retain them in normal separation mode. HILIC is a special case of normal mode separation when the high concentration of MeCN with a small amount of water is used as the mobile phase. SIELC offers a special column for HILIC separation mode: Primesep N. Primesep N is a column with a very hydrophilic surface.  It can be used for neutral polar compounds with or without buffers, offering buffer dependent selectivity and retention.