Some solutions to obtain very efficient separations in isocratic and gradient modes using small particles size and ultra-high pressure.

Details

Serval ID
serval:BIB_213DBDD36C2A
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Some solutions to obtain very efficient separations in isocratic and gradient modes using small particles size and ultra-high pressure.
Journal
Journal of chromatography. A
Author(s)
Guillarme D., Grata E., Glauser G., Wolfender J.L., Veuthey J.L., Rudaz S.
ISSN
1873-3778[electronic]
Publication state
Published
Issued date
2009
Volume
1216
Number
15
Pages
3232-43
Language
english
Abstract
The UHPLC strategy which combines sub-2 microm porous particles and ultra-high pressure (>1000 bar) was investigated considering very high resolution criteria in both isocratic and gradient modes, with mobile phase temperatures between 30 and 90 degrees C. In isocratic mode, experimental conditions to reach the maximal efficiency were determined using the kinetic plot representation for DeltaP(max)=1000 bar. It has been first confirmed that the molecular weight of the compounds (MW) was a critical parameter which should be considered in the construction of such curves. With a MW around 1000 g mol(-1), efficiencies as high as 300,000 plates could be theoretically attained using UHPLC at 30 degrees C. By limiting the column length to 450 mm, the maximal plate count was around 100,000. In gradient mode, the longest column does not provide the maximal peak capacity for a given analysis time in UHPLC. This was attributed to the fact that peak capacity is not only related to the plate number but also to column dead time. Therefore, a compromise should be found and a 150 mm column should be preferentially selected for gradient lengths up to 60 min at 30 degrees C, while the columns coupled in series (3x 150 mm) were attractive only for t(grad)>250 min. Compared to 30 degrees C, peak capacities were increased by about 20-30% for a constant gradient length at 90 degrees C and gradient time decreased by 2-fold for an identical peak capacity.
Keywords
Acetonitriles, Algorithms, Apigenin/isolation & purification, Chromatography, Liquid/methods, Coumaric Acids/isolation & purification, Kinetics, Lanatosides/isolation & purification, Models, Chemical, Molecular Weight, Pressure, Rutin/isolation & purification, Temperature
Pubmed
Web of science
Create date
04/02/2010 16:44
Last modification date
20/08/2019 12:57
Usage data