Open the Software and select the Project: “Empower “software, click “Run Sample”, and choose the project.
Edit the instrument method: Select ‘Instrument Method’, then go to “edit”
Select the pump module: Click on “W600” (the pump control).
Access the flow settings: Navigate to the “Flow” tab
Set the flow rate: For an analytical column, set the maximum flow rate to 1 ml/min to avoid over-pressurization
Adjust solvent conditions: Set the solvent composition (e.g., A% and B%). For example, in a Carboxyfantrip analysis, use 90% methanol with 0.1%TFA (A) and 10% THF (B).
Select the save method: In the method set, choose the previously saved method and click “choose”. (Note: If it’s not listed under “Develop methods”, it won’t appear; ensure it’s properly saved.)
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”).
Configure the controller: Verify the controller is directed towards the analytical setup: switch from semi prep to analytical mode.
Bypass the column for Priming: Once the method is selected, turn the lever to “R”, which diverts flow around the column and allows dripping. This will remove any air bubble. Ensure the gas is flowing.
Start solvent flow: Click “Monitor” to begin pumping solvent and flushing the system.
Adjust Flow for flushing: Press Direct on the controller to manually increase the flow rate for fast flushing.
Change flow back to 1 ml/min after bubbles are gone, then turn the lever to “C” to push through the column
Right click to customize channels: under 2D increments, uncheck e-SATIN Ch1 under 3D, check the new PAA spectrum. Apply changes, the chromatogram top will show pressure, and the bottom will display the UV vis spectrum
Once UV-Vis is leveled off, you can inject your sample. Meanwhile, you can increase the flow rate to 2 ml/min until the system is fully flushed, and any residual compounds are eluted from the column
When the chromatogram is consistent, adjust the solvent ratio back to your target composition and continue flushing.
Prepare for injection: Click the “Prepare inject button”, then inject with the lever in the “load” position.
After injection, move lever down to “inject” then click inject
After removing the syringe, return the lever to the “load” position.
Right click to customize channels → 2D increments → uncheck e-SATIN Ch1 → 3D → circle new PAA spectrum and let it run for 30-40 min. (for Carboxyfantrip, 10 minutes is fine. Peak comes out around 5:30-6:30.)
