LC Packings SWITCHOS II User Manual page 51

For a column switching experiment (a pre-concentration step) the Switchos II
Advanced Microcolumn Switching Unit, the UltiMate system (Nano configuration)
and the FAMOS Microautosampler are used and need to be programmed.
The experiment has the following features:
• NanoLC separation on a 75 µm I.D., 15 cm PepMap column.
• Injection of 10 µl of cytochrom c sample (0.1 pmol/µL).
• Sample loading onto a trap column for 5 minutes at a flow rate of 30 µL/min
(pre-concentration step).
• Solvent gradient from 0 - 50%B in 30 minutes.
• Analysis time of 60 minutes.
• Recording of the UV signal at 214 nm, the column pressure, the trap column
pressure and the pump pressure.
CHROMELEON automatically provides a program wizard when a new program is
to be created, which guides through creation of the program file.
To create a new program using the Program Wizard:
a) Select the New command on the File menu, and then select Program File from
the New box. The Program Wizard starts automatically.
b) Select the Timebase from the Select Timebase Options box.
c) Setup the column oven temperature (if applicable) in the Oven Options box.
d) Setup the trap column pressure limits of the Switchos II Loading Pump (e.g.
0 bar and 200 bar) and fill in the time table in the Loading_Pump Options box.
FIGURE 4-9
e) Setup the gradient conditions in the Pump Options box, use the 'Multi-step
Gradient' option to run a gradient.
User's Manual Switchos II
The Loading Pump Options Box
D850R3
Software Control
4-9