by
juddc » Tue Dec 08, 2009 5:39 pm
I don't know if I have ever needed to zero the pressure transducer and I don't generally pay any attention to it.
My Monday startup routine goes something like this:
1. Prepare mobile phases as needed, flush solvent lines, and prime the pump.
2. Top up the needle wash reservoir and prime the wash pump as needed. (Use a solvent that is stronger than your starting MP - I usually use neat MeOH)
3. Run the pump at a typical flow rate for a few minutes with a union in place of the column in order expel air.
4. Purge the injector for 5-10 min @ 1 ml/min.
5. Decrease the pump flow rate to ~0.1 ml/min (or whatever is an appropriate trickle flow for your column) & install column inlet, leaving the outlet open and vertical if possible to ensure constant, bubble free flow from the column outlet.
6 . Once constant flow is established at the column outlet, install the line from the column to the detector, place column in the column oven and allow it to come to desired temperature with a reduced MP flow, usually 0.5 ml/min of a 4.6mm ID column.
7. Once the column is at temperature, equilibrate for ~ 5 system volumes at a normal flow rate, longer for any ion-paired or normal phase methods.
8. Condition the column if using a gradient, re-equilibrate as needed, run the system suitability, then calibrate and quantitate with wild abandon.
Hope this helps!