Axial heterogeneities in capillary ultrahigh pressure liquid chromatography columns: Chromatographic and bed morphological characterization

Reising, Arved E.; Godinho, Justin M.; Bernzen, Janek; Jorgenson, James W.; Tallarek, Ulrich

doi:10.1016/j.chroma.2018.07.037

Cited by 10 publications

(8 citation statements)

References 53 publications

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“…It should however be noted that column packing of microparticulate columns is still highly challenging. Reising et al reconfirmed the importance of eddy dispersion on separation efficiency induced by packing heterogeneity [47,48]. Chromatographic and morphological characterizations of slurry-packed columns (at constant pressure) revealed that the most homogeneous bed microstructure and highest separation efficiency are obtained towards the column outlet [47].…”

Section: Advancing Performance Limits and Selected Examplesmentioning

confidence: 97%

“…Reising et al reconfirmed the importance of eddy dispersion on separation efficiency induced by packing heterogeneity [47,48]. Chromatographic and morphological characterizations of slurry-packed columns (at constant pressure) revealed that the most homogeneous bed microstructure and highest separation efficiency are obtained towards the column outlet [47]. In a study conducted by Billen et al, it was demonstrated that also the particle-size distribution and especially the presence of fines significantly affects the A-term contribution to chromatographic dispersion [49].…”

Section: Advancing Performance Limits and Selected Examplesmentioning

confidence: 98%

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Resolving power in liquid chromatography: A trade‐off between efficiency and analysis time

Dores-Sousa

Vos

Eeltink

2018

J of Separation Science

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This review describes chromatographic dispersion and different plate‐height models frequently used to assess the chromatographic performance of ultra‐high‐pressure liquid chromatography column technology. Furthermore, different performance indices, including the resolution, the separation impedance, and kinetic plots are discussed allowing to quantify and visualize the resolving power in liquid chromatography. The construction of kinetic plots is explained, and different visualization approaches are highlighted. Finally, key instrument and column‐technology developments to advance the kinetic performance limits are discussed and selected state‐of‐the‐art applications are highlighted.

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Section: Advancing Performance Limits and Selected Examplesmentioning

confidence: 97%

Section: Advancing Performance Limits and Selected Examplesmentioning

confidence: 98%

Resolving power in liquid chromatography: A trade‐off between efficiency and analysis time

Dores-Sousa

Vos

Eeltink

2018

J of Separation Science

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“…One explanation for this worse kinetic performance of the 50 cm column is the difficulty to pack longer columns. In particular, axial heterogeneities in the packing bed have been attributed to poor chromatographic efficiency for longer columns [38].…”

Section: Effect Of Column Length On Lipid Separationsmentioning

confidence: 99%

Ultrahigh-Performance capillary liquid chromatography-mass spectrometry at 35 kpsi for separation of lipids

Sorensen

Miller

Jorgenson

et al. 2020

Journal of Chromatography A

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Improvements in sample preparation, separation, and mass spectrometry continue to expand the coverage in LC-MS based lipidomics. While longer columns packed with smaller particles in theory give higher separation performance compared to shorter columns, the implementation of this technology above commercial limits has been sparse due to difficulties in packing long columns and successfully operating instruments at ultrahigh pressures. In this work, a liquid chromatograph that operates up to 35 kpsi was investigated for the separation and identification of lipid species from human plasma. Capillary columns between 15-50 cm long were packed with 1.7 μm BEH C18 particles and evaluated for their ability to separate lipid isomers and complex lipid extracts from human plasma. Putative lipid class identifications were assigned using accurate mass and relative retention time data of the eluting peaks. Our findings indicate that longer columns packed and operated at 35 kpsi outperform shorter columns packed and run at lower pressures in terms of peak capacity and numbers of features identified. Packing columns with relatively high concentration slurries (200 mg/mL) while sonicating the column resulted in 6 to 34 % increase in peak capacity for 50 cm columns compared to lower slurry concentrations and no sonication. For a given analysis time, 50 cm long columns operated at 35 kpsi provided a 20-95% increase in chromatographic peak capacity compared with 15 cm columns operated at 15 kpsi. Analysis times up to 4 h were evaluated, generating peak capacities up to 410 ± 5 (n = 3, measured at 4σ) and identifying 480 ± 85 lipids (n = 2). Importantly, the results also show a correlation between the peak capacity and the number of lipids identified from a human plasma extract. This correlation indicates that ionization suppression is a limiting factor in obtaining sufficient signal for identification by mass spectrometry. The result also shows that the higher resolution obtained by shallow gradients overcomes possible signal reduction due to broader, more dilute peaks in long gradients for improving detection of lipids in LC-MS. Lastly, longer columns operated at shallow

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“…It, therefore, enables a reduction of the eddy diffusion in the transchannel and short‐range intrachannel ranges. Nevertheless, maintaining all these conditions for preparing the suspension for packing in the Resing study [20] showed different column efficiencies at the column's inlet, center, and outlet. This is due to a decrease in particle–particle interactions as the packing process progresses.…”

Section: Introductionmentioning

confidence: 99%

Optimization of the packing process of microcolumns with the embedded phosphodiester stationary phases

Dembek

Szumski

Bocian

et al. 2022

J of Separation Science

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The development of new home-made stationary phases involves their packaging procedure and is crucial to obtain satisfactory working parameters. The parameter that illustrates the quality of the packed bed is its efficiency measured as the height equivalent to the theoretical plate. According to the Van Deemetr's equation, it depends on three factors, but only one of them, eddy diffusion, does not depend on the linear flow velocity. Therefore, in order to obtain it as low as possible, it is necessary to focus on a good filling of the column. Among many parameters affecting the quality of column packing, in our work we have focused on the choice of slurry solvent. Novel stationary phases with an embedded phosphodiester group were investigated. The suspensions in 16 solvents and solvent mixtures were studied for their stability, aggregation, sedimentation, and viscosity comparison. The efficiency of the packed microcolumns and its comparison was determined by chromatographic analyses using a polar (thymidine) and a nonpolar compound (naphthalene). The results obtained led to the conclusion that for these stationary phases, the best slurry solvent is the one that aggregates the phase while maintaining stability and having high viscosity.

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Axial heterogeneities in capillary ultrahigh pressure liquid chromatography columns: Chromatographic and bed morphological characterization

Cited by 10 publications

References 53 publications

Resolving power in liquid chromatography: A trade‐off between efficiency and analysis time

Resolving power in liquid chromatography: A trade‐off between efficiency and analysis time

Ultrahigh-Performance capillary liquid chromatography-mass spectrometry at 35 kpsi for separation of lipids

Optimization of the packing process of microcolumns with the embedded phosphodiester stationary phases

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Axial heterogeneities in capillary ultrahigh pressure liquid chromatography columns: Chromatographic and bed morphological characterization

Cited by 10 publications

References 53 publications

Resolving power in liquid chromatography: A trade‐off between efficiency and analysis time

Resolving power in liquid chromatography: A trade‐off between efficiency and analysis time

Ultrahigh-Performance capillary liquid chromatography-mass spectrometry at 35 kpsi for separation of lipids

Optimization of the packing process of microcolumns with the embedded phosphodiester stationary phases

Contact Info

Product

Resources

About

Ultrahigh-Performance capillary liquid chromatography-mass spectrometry at 35 kpsi for separation of lipids