1260 Infinity Binary LC - System User Guide
65
Optimization of the Agilent 1260 Infinity Binary LC
3
How to Prevent Column Blockages
How to Prevent Column Blockages
Columns packed with sub- 2- micron particles also need frits with small pore
size to prevent packing material being swept out. This immediately brings
increased risk of blocking these frits with particulates from the sample, mobile
phase or/and from the instrument itself. To protect the column, additional
small filters (
on page 66) can be used in front of the column. It is
also recommended that the sample is thoroughly filtered or/and centrifuged,
and that any kind of particulate matter in the mobile phases is avoided.
To ensure best results follow these simple usage guidance:
1
Install and run the column only in the flow direction marked on the
column.
2
Use only solvents that are high quality, chromatography grade.
3
Filter all aqueous buffers and all samples through an appropriate 0.2 µm
filter before use.
4
Replace bottles of mobile phase buffer every 24 – 48 h — do not add
mobile phase to the bottle; always use a new bottle.
5
Do not use a high buffer salt mobile phase (> 50 mM) in combination with
high acetonitrile concentrations due to possible precipitation.
6
An in- line filter is recommended to catch particulates and extend column
life. Use the suitable 1260 Infinity In- line filter for your column: 1260
Infinity LC in- line filter, 2.1 mm, 0.2 µm pore size filter, max 600 bar,
connecting capillary 70 x 0.12 mm SS (5067- 1551) for 2.1 i.d. columns,
1260 Infinity LC in- line filter, 4.6 mm, 0.2 µm pore size filter, max 600 bar,
connecting capillary 90 x 0.17 mm SS (5067- 1553) for 4.6 mm or 3.0 mm
i.d. columns. Change the filter when the pressure increases by 10 %.
7
Purge the pumps (the connections up to the column) of any buffer
containing mobile phases and flush through 5 mL of solvent before
attaching the column to the instrument.
8
Flush the column with compatible mobile phase starting slowly at
0.1 mL/min for a 2.1 mm inner diameter column, 0.2 mL/min for a
3.0 mm inner diameter column, and 0.4 mL/min for 4.6 mm inner
diameter. Increase the flow rate to the desired flow over 5 minutes.
9
Once the pressure has stabilized, attach the column to the detector.
10
Equilibrate the column and detector with 10 column volumes of the
mobile phase prior to use. (1 – 5 mL depending on column size.)
11
Avoid over- pressure. Check the pressure range of your gradient — which
may be 100 – 130 bar or more — before starting any sequence.
