NanoLC 1D Plus and 2D System Operator's Manual - Eksigent

30 Operator’s ManualFigure 4-8. Flow Calibration Windowstep 2step 3step 4step 5Verify that mobile phases specified are correct. If incorrect, click Cancel to close the Flow meterCalibration Window. Make the necessary changes in the Mobile Phases Window, perform step 1again and then click Next.Attach the flow calibration assembly, with appropriate pipette to either the mixing tee or after thevalve or column. Select the appropriate pipette size and flow rate for calibrating that channel (seetable 4-2).20 µL/division in the pipette size to set the calibration flow rate to 5 µL/min for the highflow channel or 1 µL/division in the pipette size to set the calibration flow rate to 500 nL/min for thelow flow channel.Click Next to start the flow in channel A. Enter the appropriate volume requested in step 3 of theFlow Calibration Window dialog (Fig. 4-8). Wait until the liquid front travels to the black line mark onthe pipette and press Start to begin timing. For Channel 1, time how long it takes for the liquid frontto travel 20 µL. For Channel 2, time how long it takes for the liquid front to travel 2 µL. Press Stopwhen the fluid front reaches the end of the pipette, or the appropriate mark. Then click Next. Ifnecessary disconnect the calibration assembly and dry out the liquid inside the capillary.Repeat the measurement for calibration of the B channel flow meter.step 6Verify that the instructions were followed exactly and that all values entered are correct and then clickfinish.nanoLC Operator’s Manual 611-00012 Revision A1