Evaluating Relative Retention of Polar Stationary Phases in Hydrophilic Interaction Chromatography

Guo, Yong; Bhalodia, Nidhi; Fattal, Bassel

doi:10.3390/separations6030042

Cited by 5 publications

(5 citation statements)

References 26 publications

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“…This is the case, for instance, of inorganic salts in RPLC. In HILIC these salts cannot be used because of its strong polarity and several studies propose toluene as hold-up volume marker due to its non-polar hydrophobic nature [15,20,24,[30][31][32], assuming that it is unable to penetrate into the water layer adsorbed on the surface of the bonded phase. However, this is a simplified approximation since toluene was found to be more retained in a ZIC-pHILIC column than many other compounds in acetonitrile-water and methanol water MPs [33].…”

Section: Measurement Of Hold-up Volumesmentioning

confidence: 99%

HILIC characterization: Estimation of phase volumes and composition for a zwitterionic column

Redón

Subirats

Rosés

2020

Analytica Chimica Acta

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A methodology for the estimation of the different phase volumes in HILIC is presented. For a ZIC-HILIC column the mobile phase volume (hold-up volume) is determined in several acetonitrile-and methanol-water compositions by a Linear Free Energy Relationships (LFER) homologous series approach involving n-alkyl-benzenes, -phenones, and -ketones. We demonstrate that the column works as a HILIC column when the mobile phase contains high and medium proportions of methanol or acetonitrile. However, for acetonitrile contents below 20%, or 40% for methanol, same column works in RPLC. In between, a mixed HILIC-RPLC behavior is observed, and solutes of low molecular volume are retained as in HILIC mode, but the largest ones show RPLC retention. From the homologous series retention data and pycnometric measurements involving the pure organic solvents and their mixtures with water, the mean solvent composition of the water-rich transition layers between column functionalization and the bulk mobile phase, which act as stationary phase, is estimated. Finally, the phase ratio between stationary and mobile phases is also estimated for each eluent composition, allowing the calculation of the corresponding stationary phase volumes. All volumes are strongly dependent on the water content in the eluent, especially when acetonitrile is selected as mobile phase constituent. In HILIC mode, when the water content in the hydroorganic mobile phase increases, the volumes of mobile phase decrease, but the volumes of stationary phase (mainly the water layer adsorbed onto the bonded-phase and the water-enriched interface) increase. However, at high water concentrations, where the column works in RPLC mode, the mobile phase volume increases and the stationary phase (which is now the bonded zwitterion) decrease when increasing the water percentage in the mobile phase.

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Section: Measurement Of Hold-up Volumesmentioning

confidence: 99%

HILIC characterization: Estimation of phase volumes and composition for a zwitterionic column

Redón

Subirats

Rosés

2020

Analytica Chimica Acta

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“…As already mentioned in the introduction, compounds such as benzene or toluene are commonly used as HILIC hold-up volume marker [ 5 , 11 , 18 , 40 , 41 , 42 , 43 ]. However, in a previous work it was found that these particular solutes are partially retained and larger solutes of the same type, such as octylbenzene and dodecylbenzene, should be more appropriate hold-up markers [ 24 , 35 ].…”

Section: Resultsmentioning

confidence: 99%

Evaluation of Hold-Up Volume Determination Methods and Markers in Hydrophilic Interaction Liquid Chromatography

2023

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Common methods for hold-up time and volume determination in Reversed-Phase Liquid Chromatography (RPLC) have been tested for Hydrophilic Interaction Liquid Chromatography (HILIC). A zwitterionic ZIC-HILIC column has been used for the testing. The pycnometric determination method, based on differences in column weight when filled with water or organic solvent, provides the overall volume of solvent inside the column. This includes the volume of eluent semi-sorbed on the packing of the column, which acts as the main stationary phase. The homologous series approach, based on the retention behavior of homologues in relation to their molecular volume, allows the determination of accurate hold-up volumes. However, the application of this method is time-consuming. In some cases, large neutral markers with poor dipolarity/polarizability and hydrogen bonding interactions can be used as hold-up volume markers. This is the case of dodecylbenzene and nonadecane-2-one in clearly HILIC behaving chromatographic systems, the use of decanophenone as a marker can be even extended to the boundary between HILIC and RPLC. The elution volume of the marker remains nearly unaffected by the concentration of ammonium acetate in the mobile phase up to 20 mM. The injection of pure solvents to produce minor base-line disturbance as hold-up markers is strongly discouraged, since solvent peaks are complex to interpret and depend on the ionic strength of the eluent.

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“…The crossed‐linked diol phase (LUNA HILIC column) has been found to be less retentive possibly due to the reduced number of hydroxyl groups on the polymer surface. Other polyhydroxyl phases (YMC PVA‐Sil and Epic HILIC‐HC columns) exhibit stronger retention for polar compounds (Guo, Bhalodia, & Fattal, 2019). The amide group can be found in the chemically bonded amide phase and polyacrylamide phase (TSKgel Amide 80), which are more popular than the diol phase in terms of application.…”

Section: Polar Stationary Phases For Hilicmentioning

confidence: 99%

“…Dinh et al compared the retention of 21 polar stationary phases based on the average retention factors of neutral compounds (α‐hydroxy‐γ‐butyrolactone, cytosine, dimethylformamide, thiocytosine, and uracil) in a mobile phase containing 80% acetonitrile and 25 mM ammonium acetate (Dinh et al, 2011). Guo et al evaluated the retention of 25 polar columns using cytosine as the model compound in the mobile phase containing 85 and 90% acetonitrile and 5 mM ammonium acetate (Guo, Bhalodia, & Fattal, 2019). Both studies find similar ranking of the columns, as shown in Figure 10.…”

Section: Understanding the Hilic Stationary Phasesmentioning

confidence: 99%

“…Diol phase seems to be very popular among the neutral phases in terms of column offerings probably because most column manufacturers have an existing line for NPLC. The diol phase has a moderate adsorbed water layer and retentivity for polar compounds (Guo, Bhalodia, & Fattal, 2019). A pentadiol phase with five hydroxyl groups has been developed and is expected to be more polar than the diol phase.…”

Section: Polar Stationary Phases For Hilicmentioning

confidence: 99%

See 1 more Smart Citation

A survey of polar stationary phases for hydrophilic interaction chromatography and recent progress in understanding retention and selectivity

Guo

2022

Biomedical Chromatography

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Various polar stationary phases are available for hydrophilic interaction chromatography (HILIC) and help drive continuous applications in biomedical, environmental, and pharmaceutical areas in the past decade. Although the stationary phases for HILIC have been reviewed previously, it is an appropriate time to take another look at the progresses made during the past 5 years. The current review provides an overview of the polar stationary phases commercially available for HILIC applications in an effort to assist scientists in selecting suitable columns. New types of stationary phases that were published in literature in the past 5 years are summarized and discussed. The trend in stationary phase research and development is also highlighted. Of particular interest is the experimental evidence for direct interactions of polar analytes with the ligands of the stationary phases under HILIC conditions. In addition, two different approaches have been developed to delineate the relative significance of the partitioning and adsorption mechanisms in HILIC, representing an important advancement in our understanding of the retention mechanisms in HILIC.

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Evaluating Relative Retention of Polar Stationary Phases in Hydrophilic Interaction Chromatography

Cited by 5 publications

References 26 publications

HILIC characterization: Estimation of phase volumes and composition for a zwitterionic column

HILIC characterization: Estimation of phase volumes and composition for a zwitterionic column

Evaluation of Hold-Up Volume Determination Methods and Markers in Hydrophilic Interaction Liquid Chromatography

A survey of polar stationary phases for hydrophilic interaction chromatography and recent progress in understanding retention and selectivity

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