Purification Solution - Developer's Guide 69
Calibration Procedures
6
Characterizing the column volume
16 Note the applied flow rate and injection volume.
17 Open the Sample Info window, update the run name and run the method.
18 Stop the run after the delay marker peak has been detected.
19 Important: if the marker signal in the detector is too low, increase the
injection volume or concentration of the sample.
20 Repeat twice; it is sufficient to measure runs longer than 5 min once or
twice.
21 Evaluate the data:
• Read the elution time of the peak at the apex.
• The elution volume without the column:
• Calculate an average elution time of data without the column.
• Multiply it by the applied flow rate.
• Subtract the volume of the restriction capillary if used and one half of
the injection volume.
• Final column volume
• Calculate the average elution time with the column.
• Multiply it by the applied flow rate.
• Subtract the elution volume without the column and one half of the
injection volume.
If the injection volume is close to or exceeds the volume of the sample loop, use Injection
Mode Full loop with overfill factor and set the factor to 5.
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