Hydrophilic interaction chromatography coupled to electrospray mass spectrometry for the separation of peptides and protein digests

Yang, Yanrong; Boysen, Reinhard I; Hearn, Milton Thomas William

doi:10.1016/j.chroma.2009.05.085

Cited by 50 publications

(37 citation statements)

References 25 publications

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“…Therefore, the HILIC platforms provided 62 % higher overall reproducibility for peak width. This data is in contrast with the fact that HILIC peak shapes are often distorted compared to RPLC [43].…”

Section: Analytical Platform Reproducibilitycontrasting

confidence: 70%

Cross-platform metabolic profiling: application to the aquatic model organism Lymnaea stagnalis

et al. 2015

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The freshwater pond snail Lymnaea stagnalis is used in several studies on molecular and behavioral neurobiology and ecotoxicology showing its successful application as a model organism. In the present study, a cross-platform metabolomic approach has been evaluated to characterize the organ molecular phenotypes of L. stagnalis central nervous system (CNS), digestive gland (DG), and albumen gland (AG). Two types of tissue disruption methods were evaluated of which beads beating was the preferred method. To obtain a broad picture of the hydrophilic and lipophilic metabolome, two complementary analytical platforms have been employed: liquid chromatography (LC) and gas chromatography (GC) coupled to high-resolution mass spectrometry. Furthermore, to increase the power to separate small polar metabolites, hydrophilic interaction liquid chromatography (HILIC) was applied. The analytical platform performances have been evaluated based on the metabolome coverage, number of molecular features, reproducibility, and multivariate data analysis (MVDA) clustering. This multiplatform approach is a starting point for future global metabolic profiling applications on L. stagnalis.

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Section: Analytical Platform Reproducibilitycontrasting

confidence: 70%

Cross-platform metabolic profiling: application to the aquatic model organism Lymnaea stagnalis

et al. 2015

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“…In addition, since the injection solvent used in HILIC is the same as the final solvent of many sample preparation techniques (liquid-liquid extraction, protein precipitation, and solid phase extraction), the need for evaporation and reconstitution of the extract is eliminated, thus increasing throughput of the overall assay. The interest in and practice of HILIC is anticipated to grow even more due to the increasing need to analyze highly polar molecules in all areas of science, including bioanalysis [3], proteomics [4,5], biopharmaceuticals, toxicology, drug metabolism and metabolic profiling, and other areas in the food and pharmaceutical industries [6][7][8][9]. HILIC has also been used as an orthogonal separation step in multidimensional chromatography [10,11].…”

Section: Introductionmentioning

confidence: 99%

Influence of stationary phase chemistry and mobile‐phase composition on retention, selectivity, and MS response in hydrophilic interaction chromatography

Fountain

Diehl

et al. 2010

J of Separation Science

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A comprehensive retention and selectivity characterization of several hydrophilic interaction chromatography (HILIC) stationary phases was performed with 28 test probes in order to study the influence of particle type, surface chemistry, and mobile-phase pH on chromatographic retention, selectivity, and MS response. Selectivity differences were compared for columns operated at both low and high pH, while ESI-MS was used to study the effects of mobile-phase pH on signal response. Additionally, acetone was explored as a potential alternative to ACN as the weak HILIC solvent. Moderate differences in selectivity were observed on the same column operated at different pH, mostly due to acidic compounds. In addition, the MS response increased when a high pH mobile phase was used, particularly for analytes that were ionized with negative ESI-MS. Even larger selectivity differences were observed for different stationary phases evaluated with the same mobile phase. Acetone was not a suitable replacement for ACN in routine HILIC separations due to differences in selectivity and MS response. Finally, the data from this study were used to establish guidelines for rapid HILIC method development of polar compounds, which is demonstrated with a mixture of histidine dipeptides and organophosphonate nerve agent metabolites.

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“…The growing interest in HILIC is partly due to its high solvent compatibility with MS in the electrospray ionization (ESI) mode [35,36]. One of the major fields currently under investigation is the use of HILIC as a separation tool for proteomic applications [33,[36][37][38], but it is also studied for use in mixed-mode [39][40][41] and two-dimensional liquid chromatography (2D LC) [42][43][44][45].…”

Section: Introductionmentioning

confidence: 99%

Hydrophilic properties as a new contribution for computer-aided identification of short peptides in complex mixtures

et al. 2012

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A new method to predict elementary amino acid (AA) composition of peptides (molar mass <1,000 g/mol) is described. This procedure is based on a computer-aided method using three combined analyses-reversed phase liquid chromatography (RPLC), hydrophilic interaction chromatography (HILIC) and capillary electrophoresis coupled with mass spectrometry-and using a software calculating all possible amino acid combinations from the mass of any given peptide. The complementarity between HILIC and RPLC was demonstrated. Peptide retention prediction in HILIC was successfully modelled, and the achieved prediction accuracy was as high as r²=0.97. This mathematical model, based on amino acid retention contributions and peptide length, provided the information about peptide hydrophilicity that was not redundant with its hydrophobicity. Correlations between respectively the hydrophobicity coefficients and RPLC retention time, hydrophilicity and HILIC retention time, and electrophoretic mobility and migration time were used for ranking all potential AA combinations corresponding to the given mass. The essential contribution of HILIC in this identification strategy and the need to combine the three models to significantly increase identification capabilities were both shown. Applied to an 18-standard peptide mixture, the identification procedure enabled the actual AA combination determination of the 14 di- to pentapeptides, in addition to an over 98 % reduction of possible combination numbers for the four hexapeptides. This procedure was then applied to the identification of 24 unknown peptides in a rapeseed protein hydrolysate. The effective AA composition was found for ten peptides, whereas for the 14 other peptides, the number of possible combinations was reduced by over 95 % thanks to the association of the three analyses. Finally, as a result of the information provided by the analytical techniques about peptides present in the mixture, the proposed method could become a highly valuable tool to recover bioactive peptides from undefined protein hydrolysates.

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Hydrophilic interaction chromatography coupled to electrospray mass spectrometry for the separation of peptides and protein digests

Cited by 50 publications

References 25 publications

Cross-platform metabolic profiling: application to the aquatic model organism Lymnaea stagnalis

Cross-platform metabolic profiling: application to the aquatic model organism Lymnaea stagnalis

Influence of stationary phase chemistry and mobile‐phase composition on retention, selectivity, and MS response in hydrophilic interaction chromatography

Hydrophilic properties as a new contribution for computer-aided identification of short peptides in complex mixtures

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