Purification Solution - Developer's Guide 67
Calibration Procedures
6
Characterizing the column volume
• Save the Method as
Column_Volume.
• Set the Stoptime to No Limit for all modules (infinite run time).
• Pump:
• Set Solvents: 75 % B.
• Clear the TimeTable.
• Injection volume 2 μL (important: increase the volume if the marker
signal is too low).
• If you are using the dual loop autosampler:
• Select Lower injection loop.
• Select Partial loop filling.
• Use Plugs settings as described in “Analytical Method Settings” on
page 16 of the Default Purification Method.
• UV detector:
• Thiourea: Set the Signal A wavelength to 242 nm with 4 nm bandwidth
and no reference.
• Uracil: Set the Signal A wavelength to 260 nm with 4 nm bandwidth
and no reference.
• Save the method changes.
6 Display the relevant UV wavelength profile (242 nm or 260 nm) in the Online
Plot.
7 If you are using the dual loop autosampler:
• equilibrate the upper loop:
• Switch the valve to the upper loop
• Right-click on the autosampler diagram and click Switch Valve to
Upper Loop, If the command is not available, the upper loop is
already selected.
• Apply a pump flow of 4 mL/min for 3 min.
• Switch the valve to the lower loop.
• Wash the needle:
• right-click on autosampler diagram
• Wash Needle > Flush Port15 s
• Purge the needle:
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