Understanding and manipulating the separation in hydrophilic interaction liquid chromatography

McCalley, David V.

doi:10.1016/j.chroma.2017.06.026

Cited by 208 publications

(109 citation statements)

References 108 publications

(160 reference statements)

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“…There are different types of stationary phases available for HILIC, such as bare silica, silica‐based amido‐, amino‐, carbamate‐, cyano‐, Diol‐, polydiol‐, and zwitterionic sulfobetaine columns . In the present study, an amide column was chosen due to the following considerations: It is non‐ionic (ionic interactions may lead to undesired slow chromatographic process and peak broadening), stable, and a good option for peptide retention and separation.…”

Section: Resultsmentioning

confidence: 99%

“…HILIC has been used for the retention of a wide range of polar substances such as amino acids and peptides, carbohydrates, endogenous compounds, and nucleosides . A few recent review articles provide insight into the retention mechanism, column classification, and factors that affect retention and selectivity in HILIC . Using HILIC, the purpose of the present study was to develop a comprehensive analytical method able to detect multitudinous bioactive peptides including highly hydrophilic peptides, in equine plasma and urine, which could be used for racehorse drug testing.…”

Section: Introductionmentioning

confidence: 99%

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A comprehensive approach to detecting multitudinous bioactive peptides in equine plasma and urine using hydrophilic interaction liquid chromatography coupled to high resolution mass spectrometry

Guan

You

et al. 2019

Drug Testing and Analysis

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Bioactive peptides possess pharmacological effects and can be illicitly used in sports. To deter such misuse, an untargeted method using high resolution mass spectrometry (HRMS) has been developed for comprehensive detection of multitudinous exogenous peptides in equine plasma and urine. Forty‐four peptides were extracted using mixed‐mode solid‐phase extraction (SPE) from plasma and urine, separated with a hydrophilic interaction liquid chromatography (HILIC) column, and detected on an HRMS instrument. Ammonium formate as a mobile phase additive had effects on HILIC retention and charge state distribution of the peptides. The acetonitrile percentage in the reconstitution solution affected the solubility of peptide neat standards and peptides in plasma and urine extracts differently. The stability of the peptides in plasma at ambient temperature was assessed. The limit of detection (LOD) was 10–50 pg/mL for most of the peptides in plasma, and ≤ 500 pg/mL for the remaining. LOD was 100–400 pg/mL for the majority of the analytes in urine, and ≤ 4000 pg/mL for the others. The method was used successfully to analyze incurred plasma and urine samples from research horses administered dermorphin. Even in the absence of reference standards, dermorphin metabolites (aFGYPS‐NH2, YaFG, and YaF) were identified. These results demonstrate that data generated with this method can be retrospectively reviewed for peptides that are unknown at the time of sample analysis without requiring re‐analysis of the sample. This method provides a powerful novel tool for detection of numerous bioactive peptides and their metabolites in equine plasma and urine for doping control.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A comprehensive approach to detecting multitudinous bioactive peptides in equine plasma and urine using hydrophilic interaction liquid chromatography coupled to high resolution mass spectrometry

Guan

You

et al. 2019

Drug Testing and Analysis

View full text Add to dashboard Cite

show abstract

“…However, retention of uracil in the HEA‐PEGDMA column was lower (retention factor 0.16) than observed in the CEA‐PEGDMA column (retention factor 0.35), using 90% ACN. The retention of the uracil is related to the polar groups on the surface of each stationary phase: greatest water uptake is observed for ionized stationary phases than for polar neutral phases , such as those with carboxyl and hydroxyl bonded groups, respectively. Thus, the immobilization of the water layer onto the CEA‐based stationary phase more effective than that for the HEA‐based stationary phase and more stable water layer increases the interactions between it and the analyte and, consequently, the uracil retention.…”

Section: Resultsmentioning

confidence: 99%

Preparation of an organic monolithic column based on carboxyethyl acrylate for capillary liquid chromatography

Gama

Lee

Bottoli

2018

Separation Science Plus

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In the current study, a monolithic capillary column based on carboxyethyl acrylate and polyethylene glycol dimethacrylate was successfully prepared by photo‐initiated polymerization in the presence of selected porogens. The columns were used in capillary liquid chromatographic separations in the hydrophilic interaction mode. The monolith composition was optimized by changing the type and amount of the polar monomers. Characterization by scanning electronic microscopy indicated the monolithic columns possessed a homogeneous column bed. After the optimization, the synthesized monolith showed good selectivity for the separation of acrylamides in the hydrophilic interaction mode, with chromatographic efficiency of about 45000 plates m−1 for the most retained compound.

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“…The popularity of HILIC is clearly shown in several reviews, e.g. , and original papers e.g . published in recent years.…”

Section: Introductionmentioning

confidence: 99%

“… published in recent years. However, the predominant separation mechanism, adsorption or partitioning, is still the subject of debate . This separation mechanism depends on many factors, such as stationary phase, mobile phase composition and analyte structure.…”

Section: Introductionmentioning

confidence: 99%

Chromatographic behavior of new deazapurine ribonucleosides in hydrophilic interaction liquid chromatography

2018

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The chromatographic behavior of new biogenic purine nucleosides in hydrophilic interaction liquid chromatography was examined on three different stationary phases, namely bare silica, and amide- and cyclofructan-based stationary phases. The effects of buffer concentration, pH and acetonitrile-to-aqueous-part ratio in the mobile phase on retention and peak shape were assessed. The retention coefficients and peak symmetry values substantially differed with respect to analytes´ structures, stationary phase properties and mobile phase composition. The bare silica column was unsuitable for these compounds under the chromatographic conditions tested due to very broad and asymmetrical peaks. Furthermore, the cyclofructan-based stationary phase provided almost Gaussian peak shapes of all deazapurine nucleosides under most conditions tested. Therefore, the cyclofructan-based stationary phase is the most suitable choice for the chromatographic analysis of nucleosides.

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Understanding and manipulating the separation in hydrophilic interaction liquid chromatography

Cited by 208 publications

References 108 publications

A comprehensive approach to detecting multitudinous bioactive peptides in equine plasma and urine using hydrophilic interaction liquid chromatography coupled to high resolution mass spectrometry

A comprehensive approach to detecting multitudinous bioactive peptides in equine plasma and urine using hydrophilic interaction liquid chromatography coupled to high resolution mass spectrometry

Preparation of an organic monolithic column based on carboxyethyl acrylate for capillary liquid chromatography

Chromatographic behavior of new deazapurine ribonucleosides in hydrophilic interaction liquid chromatography

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