Column Care - Agilent Technologies BioHPLC series User Manual

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Column Care

Columns with a particle size of 1.8 µm have a 0.5 µm inlet frit. Particulates
will block the column inlet frits and should be removed before the sample
is analyzed. Where this is not possible, an inline filter or guard column
should be used to protect and increase the lifetime of the analytical
column. The AdvanceBio Peptide Mapping column has a 2 µm inlet frit.
Samples that contain bioparticulate matter larger than 2 μm will plug the
inlet frit. Guard columns are recommended for use with such samples. It
is recommended that samples are filtered before injecting them onto any
column and all eluents are freshly prepared and filtered prior to use.
See www.agilent.com/chem/guards
guard columns.
Cleaning Your Column/Extending Column Life
For columns that can be backflushed (particles >1.8 µm), clean the
column in the reverse direction. Start with a stronger (less polar) solvent.
1. Disconnect column from detector and run wash solvents into a
beaker.
2. Start with your mobile phase without buffer salts (water/organic).
Run 10 to 20 column volumes through.
3. Next, use 100% organic (methanol or acetonitrile).
4. Check pressure to see if it has returned to normal; if not, then:
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