Eicom HTEC-500, User Manual

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18
HTEC-500
10. Precolumn
A precolumn is applied to the system to avoid contamination of the separation column by protein etc.
The lifetime of a separation column depends on the state of the precolumn. Generally speaking, a
precolumn can be used for approximately 100 injections of 10
μ l of blood or tissue denatured protein
samples or a few months with microdialysis samples. These lifetime approximations can change
dramatically depending on the method used to denature the proteins during sample preparation or
the precolumn condition. Please refer to the points below at all times.
10-1. Types of Precolumn Housing and Line Filters
There are two types of precolumn housings. One is 4 mm × 3 mm ID (code PC-03) and the other one is
5 mm × 4 mm ID (code PC-04). The PC-03 has number 3 engraved on it. The PC-04 has no engraved
number. The PC-03 is used with small inner diameter columns such as the SC-5ODS 2.1 mm ID, the
AC-GEL 2.0 mm ID and the CA-5ODS 2.1 mm ID. The PC-04 is used with the SC-5ODS 3.0 mmID, the
AC-GEL 4.6 mm ID and the GU-GEL 4.6 mm ID. The PC-04 is also used with the PP-ODS 4.6 mm ID and the
precolumn is positioned before the injector but not just before the separation column. This is because
there are no compatible packing materials that can be used with the PP-ODS. To protect the PP-ODS
column, it is recommended that an inline filter (code PP-LF) should be set upstream to the PP-ODS and
downstream to the injectors.
Injector
HPLC
Pump
PEEK tubing
(0.75 mm ID
×
150 cm)
Separation
Column
Mobile phase
(in a glass bottle)
Precolumn ECD
Microdialysis
Others
PEEK tubing
(0.75 mm ID
×
150 cm)
HPLC
Pump Separation
Column
Mobile phase
(in a glass bottle)
Injector Precolumn ECD
The precolumn position depends on the kind of samples being run. For microdialysis samples, it is set
between the pumps and the injector. In this case, the sample doesn’t go through the precolumn so
any of our packing material can be used. For other samples, such as plasma or tissue homogenate, a
precolumn should be set upstream of the separation column. In this case, the packing material should
be chosen according to the kind of separation column being used. If not, the shape of the peaks may
be influenced. For more information, please refer to Eicom reports.
Fig. 16 Precolumn Position