The mobile phase in HPLC strongly influences the chromatographic separation. Its preparation can be a source of trouble for a method, if not done correctly and consistently. This article provides some suggestions and practical tips for the mobile phase preparation in HPLC analysis. Read this article from Issue 10 of the Analytix Reporter, produced by Merck, to find out more.
The most important part of a chromatographic set up
is the column. But even though it provides retention,
the final separation also depends strongly on the
mobile phase (MP). The various effects offered by the
mobile phase influence the retention and differential
migration (selectivity) of the solutes through the
column. Therefore, during method development
and optimization, the separation might have to be
tuned by changing mobile phase parameters, such
as solvent type, additives (different buffers, ion pair
reagents), or operating conditions (gradient time/
steepness, temperature, flow rate). Often, problems
with chromatographic separation are related to an
incorrectly/inconsistently prepared mobile phase.
Hence, an inclination to use simpler mobile phases can
be observed for practical reasons of increased method
robustness, easier method transfer, and ease of use
(e.g. in clinical diagnostics or in process control).
Click on the button below to discover some HPLC tips and tricks for mobile phase preparation.
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