Extension of the C18 stationary phase knowledge by using the carotenoid test

Lesellier, Éric

doi:10.1002/jssc.201000239

Cited by 9 publications

(4 citation statements)

References 37 publications

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“…This may be explained by the significantly different properties of the C 18 stationary phases used for peptide separation in the first and second dimensions in our experiments. In accordance with the carotenoid test , XBridge C 18 (first dimension) and ZorbaxSB C 18 (second dimension) belong to different selectivity groups, notably ZorbaxSB C 18 has a specific shape selectivity and higher hydrophobicity in comparison with XBridge C 18 . The specific shape selectivity of ZorbaxSB C 18 could result in significant differences in peptide retention in the second dimension.…”

Section: Resultsmentioning

confidence: 63%

Multidimensional separation of tryptic peptides from human serum proteins using reversed-phase, strong cation exchange, weak anion exchange, and fused-core fluorinated stationary phases

et al. 2013

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Proteome profiling of crude serum is a challenging task due to the wide dynamic range of protein concentrations and the presence of high-abundance proteins, which cover >90% of the total protein mass in serum. Peptide fractionation on strong cation exchange, weak anion exchange in the electrostatic repulsion hydrophilic interaction chromatography (ERLIC) mode, RP C18 at pH 2.5 (low pH), fused-core fluorinated at pH 2.5, and RP C18 at pH 9.7 (high pH) stationary phases resulted in two to three times more identified proteins and three to four times more identified peptides in comparison with 1D nanoChip-LC-MS/MS quadrupole TOF analysis (45 proteins, 185 peptides). The largest number of peptides and proteins was identified after prefractionation in the ERLIC mode due to the more uniform distribution of peptides among the collected fractions and on the RP column at high pH due to the high efficiency of RP separations and the complementary selectivity of both techniques to low-pH RP chromatography. A 3D separation scheme combining ERLIC, high-pH RP, and low-pH nanoChip-LC-MS/MS for crude serum proteome profiling resulted in the identification of 208 proteins and 1088 peptides with the lowest reported concentration of 11 ng/mL for heat shock protein 74.

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Section: Resultsmentioning

confidence: 63%

Multidimensional separation of tryptic peptides from human serum proteins using reversed-phase, strong cation exchange, weak anion exchange, and fused-core fluorinated stationary phases

et al. 2013

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“…The effect of the particle on the stationary-phase properties was intensively investigated. 580 Numerous chromatographic tests are widely applied to evaluate the C18 bonded silica stationary phases, either to classify them or to better determine their properties, reflected in their chromatographic behavior. Because the carotenoid CSS was developed originally for supercritical fluid chromatography, it has been considered rational to ensure the validity of the "classification" reached under the conditions of HPLC.…”

Section: Lesellier Carotenoid Column Selection Systemmentioning

confidence: 99%

“…Phases of high loading as well as fused core silicas were studied. The effect of the particle on the stationary-phase properties was intensively investigated . Numerous chromatographic tests are widely applied to evaluate the C18 bonded silica stationary phases, either to classify them or to better determine their properties, reflected in their chromatographic behavior.…”

Section: Chromatographic Column Selection Systemsmentioning

confidence: 99%

Column Characterization and Selection Systems in Reversed-Phase High-Performance Liquid Chromatography

et al. 2019

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Reversed-phase high-performance liquid chromatography (RP-HPLC) is the most popular chromatographic mode, accounting for more than 90% of all separations. HPLC itself owes its immense popularity to it being relatively simple and inexpensive, with the equipment being reliable and easy to operate. Due to extensive automation, it can be run virtually unattended with multiple samples at various separation conditions, even by relatively low-skilled personnel. Currently, there are >600 RP-HPLC columns available to end users for purchase, some of which exhibit very large differences in selectivity and production quality. Often, two similar RP-HPLC columns are not equally suitable for the requisite separation, and to date, there is no universal RP-HPLC column covering a variety of analytes. This forces analytical laboratories to keep a multitude of diverse columns. Therefore, column selection is a crucial segment of RP-HPLC method development, especially since sample complexity is constantly increasing. Rationally choosing an appropriate column is complicated. In addition to the differences in the primary intermolecular interactions with analytes of the dispersive (London) type, individual columns can also exhibit a unique character owing to specific polar, hydrogen bond, and electron pair donor–acceptor interactions. They can also vary depending on the type of packing, amount and type of residual silanols, “end-capping”, bonding density of ligands, and pore size, among others. Consequently, the chromatographic performance of RP-HPLC systems is often considerably altered depending on the selected column. Although a wide spectrum of knowledge is available on this important subject, there is still a lack of a comprehensive review for an objective comparison and/or selection of chromatographic columns. We aim for this review to be a comprehensive, authoritative, critical, and easily readable monograph of the most relevant publications regarding column selection and characterization in RP-HPLC covering the past four decades. Future perspectives, which involve the integration of state-of-the-art molecular simulations (molecular dynamics or Monte Carlo) with minimal experiments, aimed at nearly “experiment-free” column selection methodology, are proposed.

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“…These observations yielded the development of a new chromatographic test to evaluate the properties of C18-bonded phases and to classify them [24]. It was shown that the 13-cis/all-trans-β-carotene separation factor, related to the three-dimensional organization of the stationary phase, depends on the type of the bonding (monomeric vs. polymeric), on the bonding density, and the pore size [25]. Additionally, it was shown that the retention factor of β-carotene is related to the stationary phase hydrophobicity, which is related both to the specific surface area and the bonding density.…”

Section: Fundamental Sfc Studies On Carotenoidsmentioning

confidence: 99%

Supercritical fluid chromatography for the analysis of natural dyes: From carotenoids to flavonoids

Lesellier

West

2021

J of Separation Science

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Plant-derived natural dyes are used in a variety of formulated products, from food to cosmetics and pharmaceutics. In addition to their color, they also provide some bioactivity. While they are mostly analyzed with high-performance liquid chromatography, supercritical fluid chromatography was also employed for several dye families, mostly for carotenoids and chlorophylls, and more recently for anthraquinones and flavonoids. These supercritical fluid chromatography methods are described in this review. Because the dyes have different structures and structural variations (polarity, isomers, etc.), the best chromatographic system to achieve their separation is not always the same. Hydrophobic stationary phases are preferred for the most hydrophobic dyes (chlorophylls and carotenoids) while polar stationary phases are preferred for the polar dyes (anthraquinones and flavonoids). Regarding the mobile phase composition, chlorophylls and carotenoids are best eluted with moderate proportions of cosolvent in CO 2 (about 40%), while the most polar glycosylated flavonoids require higher proportions of co-solvent and acidic additives. Because dyes are colorful, ultraviolet-visible detection is often sufficient, while mass spectrometry offers additional structural information. Furthermore, fundamental information can also be gained through chromatographic analysis of dyes: either solubility in supercritical fluids, in view of their extraction, or retention behavior providing an understanding of stationary phase properties.

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Extension of the C18 stationary phase knowledge by using the carotenoid test

Cited by 9 publications

References 37 publications

Multidimensional separation of tryptic peptides from human serum proteins using reversed-phase, strong cation exchange, weak anion exchange, and fused-core fluorinated stationary phases

Multidimensional separation of tryptic peptides from human serum proteins using reversed-phase, strong cation exchange, weak anion exchange, and fused-core fluorinated stationary phases

Column Characterization and Selection Systems in Reversed-Phase High-Performance Liquid Chromatography

Supercritical fluid chromatography for the analysis of natural dyes: From carotenoids to flavonoids

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