Principles and Practical Aspects of Preparative Liquid Chromatography

[Norm.]
4000
1.001
3000
2000
2.843
4.684
1000
0
1 2
3 4 5
Time [min]
Figure 3.19 High-volume injection with strong eluents in standard injection mode.
The chromatogram shows strongly distorted peak shapes of the compounds at 2.84
and 4.68 minutes. The solvent peak at 1.00 minute contains sample breakthrough –
as a consequence sample is lost.
Sample: 50 mg acetamidophen and 50 mg caffeine in 5000 µL DMSO
Column: Agilent ZORBAX SB C18, 21.1 x 150 mm, 5 µm
Applied gradient profile for standard injection mode:
Time Flow %A %B
0 37 93 7
0.6 37 93 7
6.0 37 78 22
6.1 37 2 98
9.0 37 2 98
9.1 37 93 7
14 37 93 7
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