A study of the re-equilibration of hydrophilic interaction columns with a focus on viability for use in two-dimensional liquid chromatography

Seidl, Claudia; Bell, David S.; Stoll, Dwight R.

doi:10.1016/j.chroma.2019.460484

Cited by 17 publications

(14 citation statements)

References 23 publications

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“…During his investigation on full and partial equilibration of HILIC phase, McCalley highlighted a repeatable equilibrium achieved after only 4.3 min [ 36 ]. The conclusions of Stoll and Seidl [ 37 ] were consistent with the obtention of high repeatable HILIC separations with very short re-equilibration times.…”

Section: Resultssupporting

confidence: 65%

An optimized LC-HRMS untargeted metabolomics workflow for multi-matrices investigations in the three-spined stickleback

et al. 2021

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Environmental metabolomics has become a growing research field to understand biological and biochemical perturbations of organisms in response to various abiotic or biotic stresses. It focuses on the comprehensive and systematic analysis of a biologic system’s metabolome. This allows the recognition of biochemical pathways impacted by a stressor, and the identification of some metabolites as biomarkers of potential perturbations occurring in a body. In this work, we describe the development and optimization of a complete reliable methodology based on liquid chromatography coupled to high resolution mass spectrometry (LC-HRMS) for untargeted metabolomics studies within a fish model species, the three-spined stickleback (Gasterosteus aculeatus). We evaluated the differences and also the complementarities between four different matrices (brain, gills, liver and whole fish) to obtain metabolome information. To this end, we optimized and compared sample preparation and the analytical method, since the type and number of metabolites detected in any matrix are closely related to these latter. For the sample preparation, a solid-liquid extraction was performed on a low quantity of whole fish, liver, brain, or gills tissues using combinations of methanol/water/heptane. Based on the numbers of features observed in LC-HRMS and on the responses of analytical standards representative of different metabolites groups (amino acids, sugars…), we discuss the influence of the nature, volume, and ratio of extraction solvents, the sample weight, and the reconstitution solvent. Moreover, the analytical conditions (LC columns, pH and additive of mobile phases and ionization modes) were also optimized so as to ensure the maximum metabolome coverages. Thus, two complementary chromatographic procedures were combined in order to cover a broader range of metabolites: a reversed phase separation (RPLC) on a C18 column followed by detection with positive ionization mode (ESI+) and a hydrophilic interaction chromatography (HILIC) on a zwitterionic column followed by detection with negative ionization mode (ESI-). This work provides information on brain, gills, liver, vs the whole body contribution to the stickleback metabolome. These information would help to guide ecotoxicological and biomonitoring studies.

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Section: Resultssupporting

confidence: 65%

An optimized LC-HRMS untargeted metabolomics workflow for multi-matrices investigations in the three-spined stickleback

et al. 2021

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“…Each of these RPLC based combination has its application scope. NP × RP [7] and RP × RP [8] systems are commonly used to separate medium and strong polar compounds, while IEX × RP [9], HILIC × RP [10] and SEC × RP [4] systems are suitable to separate strong polar compounds. According to research it has de cient in separating of weakly polar samples.…”

Section: Coupling An Nplc With Another Nplcmentioning

confidence: 99%

Development of an on-line two-dimensional normal phase liquid chromatography system for analysis of weakly polar samples by

Zhu

Xie

Liu

et al. 2022

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In this study, a novel on-line two-dimensional (2D) normal phase × normal phase liquid chromatography (NPLC × NPLC) was developed for the separation of weakly polar samples. The 2D NPLC was integrated by a unique designed solvent evaporation (SE) interface, including a 6-port 2-position valve and a 10-port 2-position valve in conjunction with two silica gel-packed enrichment columns. The enrichment columns played a pivotal role in evaporating normal phase (NP) solvent from the first-dimensional NPLC under vacuum and elevated temperature condition. The working parameters of the interface were evaluated comprehensively. To demonstrate the resolving powerful 2D system, we analyzed three natural-sourced weakly polar compounds from the extracts of toad venom, dammar resin, and propolis. To this end, we studied the effects of mobile phase combinations, the second dimensional column length for separation of the extracts from toad venom, and propolis respectively. In general, we have found that excellent separation can be obtained with diversity of NP solvent combinations and longer NPLC column (150 mm long). Moreover, compared with the generally used 2D reversed-phase liquid chromatography (RPLC × RPLC), the NPLC × NPLC method exhibited better separation orthogonality. In conclusion, the new NPLC × NPLC separation method provides potential advantages for analysis of weakly polar samples.

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“…The former approach has been most effective to utilizing reversed phase columns in the second dimension with only 2–3 column volumes required to achieve high retention time repeatability. − Hydrophilic interaction chromatography (HILIC) has not been utilized as frequently for high-speed separations in the second dimension or for ultrafast separations in general, partially due to the perceived need for lengthier re-equilibration times to achieve retention time repeatability . However, recent results on this topic have demonstrated that partial re-equilibration strategies compatible with reversed phase columns can also be implemented with HILIC columns, with re-equilibration times as short as 3 s being feasible on shorter columns typical of ultrafast methods. − …”

Section: Two-dimensional Lcmentioning

confidence: 99%

High-Throughput and Ultrafast Liquid Chromatography

et al. 2019

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A study of the re-equilibration of hydrophilic interaction columns with a focus on viability for use in two-dimensional liquid chromatography

Cited by 17 publications

References 23 publications

An optimized LC-HRMS untargeted metabolomics workflow for multi-matrices investigations in the three-spined stickleback

An optimized LC-HRMS untargeted metabolomics workflow for multi-matrices investigations in the three-spined stickleback

Development of an on-line two-dimensional normal phase liquid chromatography system for analysis of weakly polar samples by

High-Throughput and Ultrafast Liquid Chromatography

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