2D-LC User Guide 381
14 Theoretical Background
Practical Issues
• Instrumentation
It is important to use very low delay-volume-gradient pumping systems that
are able to produce high flow rates to achieve fast second dimension
gradients with only little gradient delay - like the Agilent 1290 Infinity LC.
• Columns
Total orthagonality is difficult to achieve, as there are relatively few
combinations sufficiently phase selective.
• Detection methods
Compared to mass spectrometry DAD based UV detection is faster, cheaper
and offers higher reproducibility, thus mass spectrometry offers additional
increase in peak capacity by expanding the separation space into the
MS-domain. A high sensitivity UV-detector is recommended since a dilution of
the first dimension peaks occurs in the second dimension separation – an
Agilent 1260 or 1290 Infinity Diode-Array-Detector with 60 mm flow cell is
ideal as second dimension detector.
• Data analysis
2D-LC-data are complex. Use of special software is advisable.
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