Journal
JOURNAL OF CHROMATOGRAPHY A
Volume 1218, Issue 1, Pages 64-73Publisher
ELSEVIER
DOI: 10.1016/j.chroma.2010.10.096
Keywords
Two dimensional liquid chromatography; Peak capacity; Optimization; Under-sampling effect; Poppe approach; Gradient elution
Funding
- NIH [GM 054585]
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In this work we develop a practical approach to optimization in comprehensive two dimensional liquid chromatography (LC x LC) which incorporates the important under-sampling correction and is based on the previously developed gradient implementation of the Poppe approach to optimizing peak capacity. The Poppe method allows the determination of the column length, flow rate as well as initial and final eluent compositions that maximize the peak capacity at a given gradient time. It was assumed that gradient elution is applied in both dimensions and that various practical constraints are imposed on both the initial and final mobile phase composition in the first dimension separation. It was convenient to consider four different classes of solute sets differing in their retention properties. The major finding of this study is that the under-sampling effect is very important and causes some unexpected results including the important counter-intuitive observation that under certain conditions the optimum effective LC x LC peak capacity is obtained when the first dimension is deliberately run under sub-optimal conditions. In addition, we found that the optimum sampling rate in this study is rather slower than reported in previous studies and that it increases with longer first dimension gradient times. (C) 2010 Elsevier B.V. All rights reserved.
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