Aspects of column fabrication for packed capillary electrochromatography

Angus, Peter D. A.; Demarest, Charles W.; Catalano, Tom; Stobaugh, John F.

doi:10.1016/s0021-9673(00)00529-x

Cited by 26 publications

(5 citation statements)

References 58 publications

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“…Further bed density variation can be found to depend on the kinetic energy of the particles while packing. At high particle kinetic energy, such as with low viscosity solvents and at high pressures, particles can forcibly displace already packed particles leading to a more densely packed bed [11]. Each of these factors affect the packing density which can lead to large variations in the eddy dispersion between columns packed under different conditions.…”

Section: Role Of Column Packing On Performancementioning

confidence: 99%

“…Many times different batches of particles resulting in the same nominal particle diameter, pore diameter, particle size distribution, and carbon load require modification of the packing conditions. Seemingly small changes in the packing conditions can produce significant variation in packing density and radial homogeneity and thus column performance [11,[16][17][18][19]. Furthermore, difficulty in obtaining radial homogeneity of the bed is inherent due to the use of liquid as the particle dispersant.…”

Section: Historical Challenges With Column Packingmentioning

confidence: 99%

“…These difficulties with column packing have been found to be exacerbated in moving to smaller diameter particles which are expected to yield greater efficiency [1,2,10,11,13,17]. As the particle size is decreased, the surface chemistry of the particle becomes more important due to the increase of the surface area to volume ratio [10].…”

Section: Historical Challenges With Column Packingmentioning

confidence: 99%

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1.1 μm Superficially porous particles for liquid chromatography

Blue

Jorgenson

2015

Journal of Chromatography A

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Section: Role Of Column Packing On Performancementioning

confidence: 99%

Section: Historical Challenges With Column Packingmentioning

confidence: 99%

Section: Historical Challenges With Column Packingmentioning

confidence: 99%

See 1 more Smart Citation

1.1 μm Superficially porous particles for liquid chromatography

Blue

Jorgenson

2015

Journal of Chromatography A

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“…Both of these steps require considerable experimental skills and experience in order to obtain stable columns with repeatable properties. Although packing is currently a well-established technique for the production of CEC columns, repeatability of this procedure remains problematic [12,13].…”

Section: Introductionmentioning

confidence: 99%

Recent advances in the control of morphology and surface chemistry of porous polymer‐based monolithic stationary phases and their application in CEC

2007

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This review focuses on developments in the field of polymer-based monolithic columns for CEC published in the literature since the beginning of the year 2005. The possibility of in-situ preparation as well as easy control over their porous properties and surface chemistries clearly make monolithic separation media an attractive alternative to capillary columns packed with particles. Different variables such as polymerization conditions, morphology, and surface chemistry are shown to directly affect performance of monolithic capillary columns in terms of efficiency, analysis time, and retention.

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“…In larger bore columns operated at higher pressures there is the potential for sufficient heat generation to cause unwanted dispersive processes; however with the increased surface to volume ratio of capillary column diameters of less than 250 μm this issue is avoided [23]. Importantly, packing techniques for the fabrication of long capillary columns packed with sub-2 micron columns [24, 25] have been established. Due to these various factors it is clearly advantageous to design a high PC chromatographic system based on utilization of capillary columns.…”

Section: Introductionmentioning

confidence: 99%

Profiling the Photochemical-Induced Degradation of Rat Growth Hormone with Extreme Ultra-pressure Chromatography–Mass Spectrometry Utilizing Meter-Long Microcapillary Columns Packed with Sub-2-µm Particles

et al. 2017

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In recent years protein therapeutics have seen increasing use in the therapeutic arena. As with traditional small molecule drug substances, one is obligated to ensure purity and stability of the various dosage forms. With these higher molecular weight therapeutics a common approach for analytical characterization is enzymatic digestion followed by gradient elution liquid chromatography with mass spectrometry detection to create a peptide map (bottom-up protein analysis). Due to the difficult to separate mixtures frequently encountered, there is the need for advanced chromatographic systems featuring increased resolution and/or peak capacity that can be operated in the gradient elution format. Presently we describe an extreme ultra-pressure liquid chromatography (XUPLC) system that has been implemented as an in-house add-on to a commercial ultra-pressure chromatography system. This add-on allows operation at the 38 Kspi range, accommodates the use of capillary columns in excess of one meter packed with sub-2 μm particles and can be operated in the gradient elution format. To evaluate the utility of this system, rat growth hormone was used as a model protein and was exposed to light (λ 254 nm) to create a stress environment. When enzymatic digests of control and stressed protein were analyzed with the XUPLC system using MS detection, greater than 92% peptide coverage was achieved, including the identification some peptides where pre-oxidation of Met residues had occurred, as well as chemistry specifically related to the photolysis of protein disulfide linkages. When the same samples were analyzed by commercial UPLC and compared to the XUPLC results, the utility of the increased peak capacity available with the XUPLC was apparent as previously co-eluting peaks were now well resolved. In particular one specific degradation route was identified where a pair of isobaric cis/trans diastereomerically related peptides were well resolved by XUPLC while they were unresolved by UPLC. Clearly the use of this system operating at the higher pressure regime with long capillary columns is and will be useful in continued investigations of protein stability, especially in cases where only subtle differences in the amino acid residues have occurred during degradation.

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Aspects of column fabrication for packed capillary electrochromatography

Cited by 26 publications

References 58 publications

1.1 μm Superficially porous particles for liquid chromatography

1.1 μm Superficially porous particles for liquid chromatography

Recent advances in the control of morphology and surface chemistry of porous polymer‐based monolithic stationary phases and their application in CEC

Profiling the Photochemical-Induced Degradation of Rat Growth Hormone with Extreme Ultra-pressure Chromatography–Mass Spectrometry Utilizing Meter-Long Microcapillary Columns Packed with Sub-2-µm Particles

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