Ultrahigh-Pressure Reversed-Phase Capillary Liquid Chromatography:  Isocratic and Gradient Elution Using Columns Packed with 1.0-μm Particles

MacNair, John E.; Patel, Kamlesh D.; Jorgenson, James W.

doi:10.1021/ac9807013

Cited by 375 publications

(249 citation statements)

References 13 publications

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“…second-generation system (MacNair et al, 1999). In another study performed in isocratic mode, the surge in linear velocity caused by mobile phase compression at the start of isocratic UHPLC runs (with pressures up to 6300 bar) was found to induce a 50% increase in the measured Knox C-term (Jerkovich et al, 2005).…”

Section: Chapter 1 Theory and Practice Of Uhplc And Uhplcemsmentioning

confidence: 99%

“…These columns were operated with HPLC instruments with an upper pressure limit of 400 bar and produced plate numbers of c. 25000 with a column dead time of 2e5 min. In the late 1990s, studies were reported from the groups of Jorgenson (MacNair et al, 1997;MacNair et al, 1999) and ) that made use of ultrasmall particles and high operating pressures. The first sub-2-micron particles were introduced by Agilent (Barber and Joseph, 2004) and Waters in 2004; at this time also the first commercial instrument with an operating pressure up to 1000 bar was introduced by Waters (Swartz and Murphy, 2005).…”

Section: Introductionmentioning

confidence: 99%

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Theory and Practice of UHPLC and UHPLC–MS

Guillarme

Veuthey

2017

Handbook of Advanced Chromatography /Mass Spectrometry Techniques

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Section: Chapter 1 Theory and Practice Of Uhplc And Uhplcemsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Theory and Practice of UHPLC and UHPLC–MS

Guillarme

Veuthey

2017

Handbook of Advanced Chromatography /Mass Spectrometry Techniques

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“…32,33 The group achieved a peak capacity of ∼300 over a 30 minute gradient by operating a 30 µm i.d. fused-silica capillary packed with 1 µm nonporous silica particles at 130 kpsi.…”

Section: Emerging Technologiesmentioning

confidence: 99%

“…fused-silica capillary packed with 1 µm nonporous silica particles at 130 kpsi. 32 More recently, Shen et al have intensively applied UPLC to MS-based shotgun proteomics. 34 In a 20 kpsi system, they achieved a peak capacity of >1000 by using a 2 m capillary column packed with 3 µm RP packing material.…”

Section: Emerging Technologiesmentioning

confidence: 99%

Multidimensional LC Separations in Shotgun Proteomics

2008

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Two modes of separation coupled with MS enable researchers to study complicated biological structures.Proteins are the molecular products of genes and play a central role in many biological processes. Protein expression occurs as a function of cellular and environmental conditions, and consequently proteins are expressed at different times and under different conditions. Insight is thus garnered by determining how protein expression has changed in a biological context.Over the past two decades, MS has become an important tool for the analysis of proteins. 1,2 One current method for the analysis of protein mixtures is proteolytic digestion followed by LC/MS/ MS. Once hard-to-handle proteins are converted into chemically well-behaved peptides, the approach overcomes many difficulties associated with protein mixture analysis. 3 Tandem MS has been particularly effective because the data can be directly used to identify peptides and subsequently infer which proteins (digested or undigested) are in the mixture. 4 This type of approach for the analysis of protein mixtures is often referred to as "shotgun" proteomics ( Figure 1).Because increasingly complicated biological structures are studied by tandem MS, the need for more powerful and highly resolving separation methods has grown. Consequently, the development and use of multidimensional LC (MDLC) in proteomics has thrived over the past few years. MDLC combines two or more forms of LC to increase the peak capacity, and thus the resolving power, of separations to better fractionate peptides prior to entering the mass spectrometer.High-resolution separation serves two functions in combination with MS. It better resolves peptides differing in charge and hydrophobicity to minimize ion suppression and improve ionization efficiency, and it simplifies the complexity of peptide ions entering the mass spectrometer to minimize undersampling. This last aspect is important because the tandem MS process is driven by data-dependent data acquisition and has a finite cycle time. A higher peak capacity and better resolving power improve the acquisition of data and can lead to a better representation of the proteins in the mixture. Improved resolution also results in a larger dynamic range.Although MDLC is by no means a new concept and has a long history, it has been enjoying a renaissance in proteomics. 5 In this article, we will provide an overview of the background, important applications, and future prospects for MDLC; in particular, we will focus on applications involving peptides. (To listen to a podcast about this feature, please go to the Analytical Chemistry website at pubs.acs.org/ac.)

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“…One is the development of an ultra-fast HPLC system consisting of a column packed with super-fine full-porous spherical particles and a special pump apparatus for delivering the mobile-phase solvent into the column under high-pressure conditions. [2][3][4] However, this approach is accompanied by new problems because the separation conditions are remarkably changed from those for conventional HPLC. Chromatography is carried out under much a faster flow velocity, hence, under extremely higher pressure conditions.…”

Section: Introductionmentioning

confidence: 99%

Moment Equations for Chromatography Using Superficially Porous Spherical Particles

Miyabe

2011

ANAL. SCI.

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IntroductionHigh-performance liquid chromatography (HPLC) has been extensively used as one of the most powerful separation techniques in the field of fine chemistry and biological science. 1 Still, however, it is now strongly required to develop a faster separation system with a high throughput. There are two major approaches for achieving the demand of the times. One is the development of an ultra-fast HPLC system consisting of a column packed with super-fine full-porous spherical particles and a special pump apparatus for delivering the mobile-phase solvent into the column under high-pressure conditions. [2][3][4] However, this approach is accompanied by new problems because the separation conditions are remarkably changed from those for conventional HPLC. Chromatography is carried out under much a faster flow velocity, hence, under extremely higher pressure conditions. The friction of the mobile phase against the packed bed of the super-fine particles leads to the generation of heat. The resulting temperature distribution taking place in both the axial and radial directions of the column provides an unfavorable influence on the peak profile, and hence on the column efficiency. [5][6][7][8] There would be a paradoxical limitation in this approach. Although the separation speed can be amplified with an increase in the mobile-phase flow velocity, the influence of the heterogeneous temperature distribution on the column efficiency must be emphasized. This is an additional problem of the ultra-fast HPLC technique, which can substantially be neglected under conventional HPLC conditions.An alternative is the development of new types of separation media, such as monoliths [9][10][11] and shell-type spherical particles. [12][13][14][15] Compared with the ultra-fast HPLC system described above, the new separation media show a different chromatographic behavior because their structural characteristics, i.e., shapes and porosities, are extremely different from those of the super-fine full-porous spherical particles. For example, in the case of shell-type particles, which consist of an inert center core and a superficial porous shell layer, the following three points can be picked up as their advantageous characteristics. At first, the whole particle size is relatively large. This is effective for preventing any unnecessary elevation of the column back pressure under high flow rate conditions. The size of interparticulate channels in a column packed with spherical particles depends on the particle diameter and their packing density. It is usually estimated to be about one third of the particle diameter, 16 and cannot independently be controlled. Second, the thickness of the superficial porous shell layer is sufficiently thin. It has been well recognized that the column efficiency under high flow-rate conditions primarily rests on the mass-transfer resistance in the stationary phase and that the reduction in the diffusion distance of sample molecules is effective for suppressing band broadening due to intra-stationary phas...

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Ultrahigh-Pressure Reversed-Phase Capillary Liquid Chromatography: Isocratic and Gradient Elution Using Columns Packed with 1.0-μm Particles

Cited by 375 publications

References 13 publications

Theory and Practice of UHPLC and UHPLC–MS

Theory and Practice of UHPLC and UHPLC–MS

Multidimensional LC Separations in Shotgun Proteomics

Moment Equations for Chromatography Using Superficially Porous Spherical Particles

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