4.7: Running a Sequence
- Page ID
- 401615
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- In the event that the pump and detector are off, select System On from the Instrument menu. The pump will begin to operate and the detector lamp will light.
- Make sure that all solvent resevoirs (top of instrument) are at least half full. If not, ask your instructors to show you how to refill the bottles.
- Make sure the waste bottle has space for at least 1 L of waste. If not, ask your instructors to remove the excess waste.
- Make sure that the waste solvent lines from the instrument are inserted all the way to the bottom of the 4L brown waste bottle on the floor.
- Allow the column to equilibrate to the first mobile phase (eg 60/40 methanol/water) before beginning the analysis. When a method is first loaded, the method parameters are immediately sent to the instrument. You can consider the system equilibrated when the column pressure stabilizes. Consult your TA.
- Within several minutes, a flat baseline should be achieved in the online plot window. Even using the most viscous mobile phase (50/50 acetonitrile/water), the operating pressure (displayed on the pump module and in the pump window on the screen) should not exceed 150 bar. If you have any difficulties achieving a flat baseline or, if the pressure exceeds 150 bar - consult your TA immediately. If flow is erratic or air bubbles are visible in the Teflon lines, your TA will assist you in re-purging the system.
- Set the post-run sequence command: Go to Sequence --> Sequence Parameters. Find the Shutdown options and choose the most appropriate option for post-run macros.
- If you are submitting just a few injections during the lab period, and you will be present at the end of the sequence, deselect (uncheck) the box for Post-sequence command/macro.
- If you are running a longer sequence to run (eg overnight), please add a checkmark to Post-sequence command/macro and select Turn Instrument Standby in the drop down menu (See Figure \(\PageIndex{1}\)).
- Press OK to save the Sequence Parameters.
Figure \(\PageIndex{1}\): Copy and Paste Caption here. (Copyright; author via source)
- Go to Sequence --> Save Sequence Template As and give your sequence template a file name.
- To begin the sequence, go to the Instrument Control Tab and press the button with the "play" icon (triangle inside a blue circle). See Figure \(\PageIndex{2}\). Your sequence will begin - watch the sample injection system to ensure the first sample is injected.
An alternative option for starting a sequence is to use the Run Control menu from the top level toolbar, and then Select Run Sequence.
5. The chromatogram in progress will be displayed in the upper left box on the screen. You may change the attenuation of the signal and the time scale by clicking on the up and down arrows on the left side of the signal window. Remember this is done only for your viewing convenience. The final chromatographic output will be automatically scaled.
6. When each analysis is complete, the output will be sent to the screen, a file, and/or the printer as specified in your method. The text report of peak retention times and integrated areas will also be output as specified in the method.