Open “Empower”, click “Run Sample” and select project.
Select ‘Instrument Method’ go to “edit”
Click “W600”
Click “Flow” tab
For analytical column max is 1 ml/min
Adjust solvent conditions A% and B%
Save condition method as “something” then close window
Select the previously saved method in the method set and click “choose” (if it is not in the method set it will not be under “develop method”).
Make sure the controller is pointed towards analytical: Semiprep →Analytical
Once you have chosen a method turn lever towards “R” which bypasses column and will drip -this will get out air bubble, make sure gas is flowing too.
Now click monitor to make it start sucking up solvent and flushing the system
You can press “direct” on the controller and change the flow rate directly (increase for flush)
Change flow back to 1 ml/min after bubbles are gone, then turn lever facing “C” to push through column
Right click → customize channels → 2D increments → uncircle e-SATIN Ch1 → 3D → circle new PAA spectrum now apply chromatogram top gives you the pressure and bottom you UV vis spectrum
Once UV-Vis is leveled off you can inject your sample. In the meantime you can increase the flow rate to 2 ml/min until everything is flushed, compounds on column
Once the chromatogram is consistent you can change the solvent ratio back to desired one and flush the system with it.
Now hit prepare inject button, then inject with the lever in the the “load” position.
After injection, move lever down to “inject” then click inject
After pulling out the syringe turn the lever back to the “load” position.
Now right click customize channels → 2D increments → uncircle e-SATIN Ch1 → 3D → circle new PAA spectrum, and let it run for 30-40 min.
