Transferring Existing HPLC Methods to UPLC™: A Systematic Approach
Ultra Performance LC™ is an analytical tool that has resulted
from the evolution of column chemistry and instrumentation. This
technique can be used to develop new analytical methods, but it also
finds utility in improving the speed, sensitivity, and resolution of
existing methods.To transfer an existing HPLC method to a highly
efficient UPLC method, the properties of the instrument, operating
conditions and the column chemistry must be controlled and matched.
Geometric considerations and scaling will usually minimize any impact
of the instrument and the procedures. The subtleties of column
chemistry have a less predictable effect on the transfer. It is
necessary to properly characterize the selectivity relationships
between UPLC columns and common HPLC materials to best match retention
and selectivity. Final adjustment of mobile phase and operating
conditions is often required for particular combinations of sample and
column in an established method.