72 Purification Solution - Developer's Guide
6
Calibration Procedures
System Configuration and Delay Time Calibration
The Purification software offers the function of a scale-up from the
analytical to the preparative flow or back using parameters of analytical
and preparative columns. Since the results of the FC delay calibration are
required for the initial setup of the software, it is easier to do a manual flow
scale-up using one of following equations:
• For columns of the same porosity:
• For columns of different porosity:
where
F = flow
d = diameter
L = column length
V
0
= column void volume
subscripts:
p
is particle,
anal
is analytical and
prep
is preparative
Porosity, in the context of the Purification software, is ratio of the void
column volume to the geometrical column volume.
• Make-up flow
Tubing design is optimized for 1.5 mL/min make-up flow that is split
approximately 1:5 by a T-junction in front of the MSD in order to have the
final flow to the MSD close to 0.2 – 0.3 mL/min. Note that decreasing the
make-up flow increases the MSD-to-FC delay; increasing it decreases the
signal response in all detectors after the splitter.
• MSD cycle time if MSD is used
The MSD cycle time is a decisive parameter in ensuring that the calibrated
delays for UV to FC and UV to MSD are sufficient. For a default setting
follow the Peakwidth recommendation in the “Preparing Default
Purification Methods” on page 9, and consider further optimization of the
value as suggested in “Insufficient MSD-to-FC delay time” on page 81.
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