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.
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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 canbe 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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