Dionex UltiMate 3000, Установка системы и приложений

Страница: 43 / 73

UltiMate 3000 Nano and Cap - System Installation and Application
39
3.6 Parallel LC
This application allows you to increase the throughput for peptide separations. While the
autosampler's injection valve is in
Load position, sample is drawn by the syringe through the
needle and placed in the sample loop. The needle pair withdraws liquid from the well or vial
and then descends into the wash port. The injection valve is switched to the
Inject position,
allowing the loading pump to direct the sample from the sample loop onto trap column 1 (in
the FLM-3200;
→
Fig. 21, no. 9) for sample preconcentration. (For information about how to
install the trap columns, refer to section 3.7.)
Valve 1 (left switching valve in the FLM-3200) is in position 10-1 . In parallel, valve 2 (in the
FLM-3300) is in position 1-2 to allow the wash step to elute remaining hydrophobic compounds
from trap column 2 (in the FLM-3300;
→
Fig. 21, no. 11) and separation column 2 (in the
FLM-3300;
→
Fig. 21, no. 19). The nano valve (right switching valve in the FLM-3200) is in
position 6-1 to direct the eluent from the wash-step to the detector.
When the sample is loaded on trap column 1, valve 1 is switched to position 1-2 . The analytical
pump 1 then directs the sample from trap column 1 to separation column 1 (in the FLM-3200;
→
Fig. 21, no. 14). At the time when the first peaks are expected, the nano valve is switched to
position 1-2 to direct the eluent to the detector. In the meantime, separation column 2 is
reconditioned. Shortly before the separation ends, valve 2 is switched to position 10-1 . Trap
column 2 is conditioned with the loading solvent.
As soon as the gradient over separation column 1 is finished, a new sample is injected. Valve 1 is
in position
1-2. Valve 2 is in position 10-1 to load the sample on trap column 2. At the same time,
separation column 1 is washed and its eluent is collected, with the nano valve being in position 1-
2 .
When the trap column 2 is loaded, valve 2 is switched to position 1-2 . This connects the
separation column 2 with the trap column 2 (analytical pump). At the end of the wash step for
separation column 1, the nano valve is switched to position
6-1 . The separation column 1 is
reconditioned while the separation runs on separation column 2. Before the experiment ends,
valve 1 is switched to position
10-1 to prepare for the next injection.
This sequence, which can be repeated as often as needed, allows running the analysis on one
column and simultaneously washing and equilibrating the other column.
Please note: The application comprises two pumps (LPG-3400 and DGP-3600) and two
flow managers (FLM-3200 and FLM-3300;
→
Fig. 20, page 40).
Therefore, adapt the names under which the devices are identified in the
installation environment and in the Chromeleon client in the Chromeleon
Server Configuration program. Keep in mind that you may have to adapt
the links to the associated control panels as well.