HPLC also called high-pressure liquid chromatography, is
a chromatographic separation technique used in analytical chemistry to identify
and quantify the individual analytes of sample solutions. The method
development in HPLC needs optimization of different types of mobile phase and
column parameters, the pH of the mobile phase being one of its examples. In the
reversed-phase HPLC, for the separation of acid and base samples, the pH of the
mobile phase plays a significant role in determining selectivity and, retention
of a molecule, and in managing the ruggedness and reproducibility of a method.
Considerate how the pH of the mobile phase influence your
HPLC columns and components can assist you quickly conclude the suitable mobile
phase conditions for a separation, decreasing the time it takes to develop a
rugged HPLC method. The mobile phase pH can affect the peak shape and retention
time of a molecule since it affects the ionization state of the molecule, and
hence the chemistry of interactions happening within the stationary phase. The
acidic component has increased retention in low pH and reduced retention in
high pH. Since at low pH bases are ionized and poorly retained, but at high pH,
they are neutral and well retained.
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