Purification Solution - Developer's Guide 59
Calibration Procedures
6
Characterizing the delay volumes: Mixing point to UV detector and Column to UV detector
• Save the Method as
Prep_Mixing_Point_To_UV.
• Set Stoptime to No Limit for all modules (infinite run time).
• For the combined instrument, set the 2/10 port valve in the method to
the preparative flow path.
• Dual-loop autosampler if used:
• Set the Injection loop: Upper
• Make sure that the upper loop is in the main pass. Right-click on the
autosampler diagram and check if the command Switch Valve to Upper
or Lower Loop refers to the lower loop; if not, change to the upper loop
using this command.
• Set the UV detector wavelength to 263 nm with 4 nm bandwidth and
without a reference.
• Prep Pump Cluster:
• Set the flow to 2 mL/min.
• Set the solvent composition to 40 % B.
• In the Advanced Channel A section, set the Compressibility to 46.
• In the Advanced Channel B section, set the Compressibility to 46.
• Set following pump time table:
• Save the method changes.
5 Display the 263 nm UV profile (signal A) in the Online Plot.
6 Switch on all modules.
7 Equilibrate the system with 40 % solvent B at 2mL/min for 5min.
8 If the pressure is below 20 bar, place a restriction capillary of known
volume (0.12 × 2000 mm) in the position of the column and equilibrate the
system again.
Time [min] A [%] B [%] Flow [mL/min] Max. Pressure Limit [bar]
0.00 60.0 40.0 2.000 200.00
1.99 60.0 40.0 --- ---
2.00 40.0 60.0 --- ---
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