Comparison of the quantitative performance of constant pressure versus constant flow rate gradient elution separations using concentration-sensitive detectors
Autor: | Gert Desmet, Pat Sandra, Klaus Witt, Matthias Verstraeten, Konstantin Choikhet, Ken Broeckhoven, Frederic Lynen, Monika Dittmann |
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Přispěvatelé: | Chemical Engineering and Industrial Chemistry, Chemical Engineering and Separation Science |
Rok vydání: | 2012 |
Předmět: |
Analytical chemistry
Wine xx Biochemistry High-performance liquid chromatography Antioxidants Analytical Chemistry Viscosity Limit of Detection Phase (matter) Pressure Calibration Animals Chromatography High Pressure Liquid Chromatography Chemistry Elution Organic Chemistry Reproducibility of Results Linearity Serum Albumin Bovine General Medicine Repeatability Peptide Fragments Volumetric flow rate Models Chemical Linear Models Cattle Steroids |
Zdroj: | Vrije Universiteit Brussel |
ISSN: | 0021-9673 |
DOI: | 10.1016/j.chroma.2011.10.019 |
Popis: | This contribution discusses the difference in chromatographic performance when switching from the customary employed constant flow rate gradient elution mode to the recently re-introduced constant pressure gradient elution mode. In this mode, the inlet pressure is maintained at a set value even when the mobile phase viscosity becomes lower than the maximum mobile phase viscosity encountered during the gradient program. This leads to a higher average flow rate compared to the constant flow rate mode and results in a shorter analysis time. When both modes carry out the same mobile phase gradient program in volumetric units, normally identical selectivities are obtained. However, small deviations in selectivity are found due to the differences in pressure and viscous heating effects. These selectivity differences are of the same type as those observed when switching from HPLC to UHPLC and are inevitable when speeding up the analysis by applying a higher pressure. It was also found that, when using concentration-sensitive detectors, the constant pressure elution mode leads to identical peak areas as the constant flow rate mode. Also the linearity is maintained. In addition, the repeatability of the peak area and retention time remains the same when switching between both elution modes. |
Databáze: | OpenAIRE |
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