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Minimum retention at the "U turn" composition of the mobile phase corresponds to the transition from the RP to the NP (HILIC in an aqueous organic mobile phase) mechanism, and the retention at this point is usually very low [ 94, 99].
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The next two chapters deal with different general parameters influencing modelling of the retention mechanism (mobile phase composition and nature of the analyte).
It may be concluded that for low pH (4.0) of the mobile phase buffer, the retention mechanism of nucleotides is of mixed nature.
A high content of acetonitrile (>75%) and low concentration of formic acid in the mobile phase promote the hydrophilic separation mechanism for MPAEs on the amide SP. So, this dual separation mechanism can be modulated modifying the pH and content of organic modifier in the mobile phase, allowing greater flexibility to develop improved methods.
The parameter m1 characterizes the effect of increasing the water concentration in the mobile phase on the contribution of the RP mechanism to the retention, whereas the parameter m2 is a measure of the opposite effect, characteristic of the HILIC contribution to the retention in highly organic mobile phases.
The characteristics of the hydrophilic stationary phase may affect and in some cases limit the choices of mobile phase composition, ion strength or buffer pH value available, since mechanisms other than hydrophilic partitioning could potentially occur.
Retention mechanism switching was easily realized by adjustment of the mobile phase constitution.
However, the participation of phenyl groups in separation mechanism decreases with the increase of acetonitrile in the mobile phase.
-Catechin and -epicatechin are separated by a reverse phase mechanism on a C18 column with an acidic acetonitrile-water mobile phase gradient.
A typical hydrophilic interaction chromatography mechanism was observed when the content of acetonitrile in the mobile phase was higher than 20%.
A typical hydrophilic interaction chromatography mechanism could be observed when the content of acetonitrile (ACN) in the mobile phase exceeded 25%.
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