Eric Chan on October 4, 2023 at 1:09 am Hello Alexandra, I’ve just started using the OptiFlow Turbo V with the nano add-on on our 7600. However, I’ve noticed even with 50 fmol on-column (350nL/min, nano gas = 15, CUR = 30, nano source temperature = 300), only some of the peptides are detectable based on XICs. Could you share with us how much of the PepCalMix you typically load on-column (with 1uL injection) for calibration? It would be great if you can share with us the source parameters (Nano gas, Curtain gas, and Nano cell temperature) as well as LC parameters (I’ve noticed you’re flowing from 5% to 30% B, but what flow rate do you run at?) They would serve as great reference points as I try to optimize the method for my system. Thanks for your help. Log in to Reply
Christie Hunter on December 1, 2023 at 5:58 pm Hi Eric, Our typical loading for nanoLC is 20fmol of PepCalMix on column, that should give you lots of signal to calibrate on. Typically we use a flow rate of 300-400nL/min depending on the column used. For source conditions, typical settings for nanoflow will be in these ranges: GS1 – 10-20 ISV – 2800 – 3000V TEM – 150-225C CUR – 25-35 Are you generating the XICs in Explorer and how recently have you calibrated the instrument? One trick to try is to increase the XIC width to 0.5 Da and see if you can now see the peaks. You might need to calibrate with the CDS and the TurboV source before starting nanoLC if you calibration is off by too much. The automatic calibration routines will only work when your calibration is off by a smaller amount (say 100 ppm). Hope this helps! Log in to Reply
Eric Chan on October 9, 2023 at 3:50 pm In case anybody is in the same situation, with our 7600 paired with an M-Class, we were unable to populate the Rack/Vial locations in SCIEX OS 3.1.6. However, all is well once we’ve upgraded to SCIEX OS 3.3. 1 Log in to Reply