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How long should the HPLC column be equilibrated?

Equilibration time is dependent on the flow rate and dimension of the column. In general, flushing the column with ten up to twenty column volumes is enough for equilibration. Therefore the flow rate has to be kept in mind. It is possible to increase the flow rate during equilibration to shorten the equilibration time. If a column should be equilibrated with a buffered mobile phase or with a mobile phase containing an ion pair reagent, the equilibration time should be extended. Using the KNAUER Method Converter you can calculate the equilibration time for your column in consideration of flow rate, column dimension, particle size and even gradient steps.

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Based in Berlin, KNAUER has been serving the sciences since 1962. We develop and manufacture scientific instruments of superior quality for liquid chromatography systems and components, including:

Analytical HPLC / UHPLC
Preparative HPLC
Fast protein liquid chromatography (FPLC)
Multi-Column Chromatography / SMB
Osmometry

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