Mobile phases in HPLC
Mobile phase is the most important parameter in reversed-phase HPLC. Type of mobile phase
used may have a big effect on the retention. It can promote or suppress an ionization of the
analyte molecules, and it also can shield an accessible residual silanol or any other active
adsorption centers on the adsorbent surface.
Proper selection of the mobile phase is the second most important step in the development of the
separation method (the first one is the selection of the adsorbent type). The main requirement for
the mobile phase is that it has to dissolve the analytes up to the concentration suitable for the
detection. Variation of the eluent composition provides the great flexibility of HPLC separations.
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