Effect of first dimension phase selectivity in online comprehensive two dimensional liquid chromatography (LC×LC)

Gu, Haiwei; Huang, Yuan; Filgueira, Marcelo R.; Carr, Peter W.

doi:10.1016/j.chroma.2011.07.063

Cited by 18 publications

(14 citation statements)

References 54 publications

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“…The chromatograms corresponding to online LC×LC runs, where gradient elution was used in both 1 D and 2 D, are explained in detail in our previous work 9 . A general description of the conditions is provided here:…”

Section: Methodsmentioning

confidence: 99%

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A Simple, Robust Orthogonal Background Correction Method for Two-Dimensional Liquid Chromatography

2012

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Background correction is a very important step that must be done before peak detection or any quantification procedure. When successful, this step greatly simplifies such procedures and enhances the accuracy of quantification. In the past, much effort has been invested to correct drifting baseline in one dimensional chromatography. In fast online comprehensive twodimensional liquid chromatography (LC×LC) coupled with diode array detector (DAD), the change in the refractive index (RI) of the mobile phase in very fast gradients causes extremely serious baseline disturbances. The method reported here can be combined with many of existing baseline correction methods for one dimensional (1D) chromatography in two dimensional (2D) liquid chromatography and recreate the background structure for further correction. When such methods are applied orthogonally to the second dimension ( 2 D), the background correction is dramatically improved. It results in an almost zero mean background level and it provides better background correction than does a simple subtraction of a blank. Indeed, the method proposed does not require running a blank sample.

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Section: Methodsmentioning

confidence: 99%

“…The experimental procedure for sample preparation and system configuration is described in detail in the given reference 9 .…”

Section: Methodsmentioning

confidence: 99%

A Simple, Robust Orthogonal Background Correction Method for Two-Dimensional Liquid Chromatography

2012

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“…Their approach shows that, due to their high retentivity and hydrophobicity, carbon‐clad columns provide rather different selectivities than the vast majority of alkyl bonded RP columns. The conclusion of Zhang et al that these columns should be used as the 2 D column in RPLC × RPLC systems has recently been adopted by Gu et al []. Jandera and coworkers used linear‐free‐energy relationships (LFER) [] as a tool for comparing the selectivity of several LC columns [].…”

Section: Quality Descriptors In Lc × Lc Optimization: the Role Of Peamentioning

confidence: 99%

Theories to support method development in comprehensive two‐dimensional liquid chromatography – A review

Bedani

Schoenmakers

Janssen

2012

J of Separation Science

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On-line comprehensive two-dimensional liquid chromatography techniques promise to resolve samples that current one-dimensional liquid chromatography methods cannot adequately deal with. To make full use of the potential of two-dimensional liquid chromatography, optimization is required. Optimization of two-dimensional liquid chromatography is a relatively new yet important research topic the aim of which is to predict combinations of stationary and mobile phases, column formats, and chromatographic conditions that maximize resolving power and minimize analysis time. In on-line two-dimensional liquid chromatography, dilution-related issues play also an important role and these should be taken into account when developing optimization strategies. In this work, state-of-the-art strategies that support method development for on-line two-dimensional liquid chromatography through a rigorous choice of chromatographic parameters are critically reviewed. The final aim is to provide practitioners with a clear understanding of which aspects can be optimized using current on-line two-dimensional liquid chromatography strategies (and which ones cannot). In two-dimensional liquid chromatography, maximizing resolving power for a given analysis time and dilution requires optimizing efficiency, selectivity and retention. While great strides forward have been made in the optimization of efficiency-related issues, considerable effort needs still to be made in terms of (1) developing models that can predict the retention factors that given stationary/mobile phase systems can provide and (2) using this information for choosing the two ones that maximize two-dimensional liquid chromatography orthogonality. Because of this limitation, in two-dimensional liquid chromatography, this aspect is typically dealt with a posteriori through examining chromatograms. This review clearly shows that important progress in the optimization of on-line two-dimensional liquid chromatography has recently been made.

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“…Mentioned combinations of different separations provide high separation orthogonality and yield increased number of identified proteins . The combination of HILIC‐RPLC has shown that equal or even higher orthogonality can be achieved compared to the most commonly employed SCX‐RPLC systems and the peptide separation combination of high pH RP and RP at low pH .…”

Section: Introductionmentioning

confidence: 99%

Direct injection of HILIC fractions on the reversed‐phase trap column improves protein identification rates for salivary proteins

2016

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Online combination of hydrophilic interaction chromatography (HILIC) and RP chromatography for separation of tryptic peptides is a challenging approach due to the incompatibility of direct loading HILIC fractions on the RP trapping column. High amounts of organic modifiers in loading solvents decrease the binding efficiency of tryptic peptides on C18 phases and lower the number of identifications. A 500 μL loop upfront of the trapping column filled with aqueous mobile phase was employed as a mixing chamber and enabled direct injections and improved saliva protein identification rates of HILIC fractions.

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Effect of first dimension phase selectivity in online comprehensive two dimensional liquid chromatography (LC×LC)

Cited by 18 publications

References 54 publications

A Simple, Robust Orthogonal Background Correction Method for Two-Dimensional Liquid Chromatography

A Simple, Robust Orthogonal Background Correction Method for Two-Dimensional Liquid Chromatography

Theories to support method development in comprehensive two‐dimensional liquid chromatography – A review

Direct injection of HILIC fractions on the reversed‐phase trap column improves protein identification rates for salivary proteins

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