Enhanced fluidity liquid chromatography for hydrophilic interaction separation of nucleosides

Treadway, James W.; Philibert, Gwenaelle S.; Olesik, Susan V.

doi:10.1016/j.chroma.2010.12.059

Cited by 30 publications

(9 citation statements)

References 26 publications

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“…As previously discussed 17 , efficiency in EFLC with HILIC does not follow the increase observed in many other applications of EFLC 11,13,14,15 which can be explained by the significant increase in retention with CO 2 addition. As seen in Figure 7, the efficiency of the monophosphates, which saw a great increase in retention factor, decreases of 35% in average, while the variation in most nucleosides' efficiency is not significant.…”

Section: Impact Of Co 2 Additionmentioning

confidence: 54%

“…We formerly established that the separation of the nucleosides was faster and more efficient that previous isocratic separation including our previous EFLC study. 17 When comparing the separation of monophosphate nucleotides to other HILIC studies, 22,23,24 the resolution and selectivity obtained with HILIC-EFLC was either comparable or superior. The best and fastest separation of monophosphate nucleotides in HILIC was achieved by Zhou et al 24 in 7 minutes at 1ml/min.…”

Section: Comparison To Other Hilic Separationsmentioning

confidence: 99%

“…31 Our previous study on hydrophilic interaction-EFLC showed satisfactory results using acetate buffers for the separation of RNA nucleosides. 17 For this study, both sodium acetate and ammonium acetate buffers were considered with concentration varying from 20 to 200mM but all led to long retention times with broadened and tailed peaks for the nucleotides. Phosphate buffers were chosen as possible alternatives because previous studies have shown value in their use in the separation of biomolecules by HILIC.…”

Section: Choice Of Buffermentioning

confidence: 99%

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Characterization of enhanced-fluidity liquid hydrophilic interaction chromatography for the separation of nucleosides and nucleotides

Philibert

Olesik

2011

Journal of Chromatography A

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Hydrophilic Interaction Chromatography (HILIC) is a liquid chromatographic separation mechanism commonly used for polar biological molecules. While nucleotides are very polar analytes, only a few studies have been conducted on their separation using HILIC. Herein, the use of enhanced-fluidity liquid chromatography (EFLC) for the separation of nucleosides and nucleotides under HILIC conditions is investigated. Enhanced-fluidity liquid chromatography involves using common mobile phases with the addition of substantial proportions of a dissolved gas which provides enhanced mobile phase diffusivity and lower viscosity. The impact of varying mobile phase composition: buffer composition, type of base, salt used, salt concentration and mole fraction of CO 2 was studied to provide optimized HILIC separations. Each of these parameters plays a key role in the retention of the analytes, which demonstrates the complexity of the retention mechanism in HILIC. The tailing of phosphorylated compounds was overcome with the use of phosphate buffer and the addition of a strong base; efficiency and peak asymmetry were compared with the addition of either triethylamine (TEA), 1,4-diazabicyclo

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Section: Impact Of Co 2 Additionmentioning

confidence: 54%

Section: Comparison To Other Hilic Separationsmentioning

confidence: 99%

Section: Choice Of Buffermentioning

confidence: 99%

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Characterization of enhanced-fluidity liquid hydrophilic interaction chromatography for the separation of nucleosides and nucleotides

Philibert

Olesik

2011

Journal of Chromatography A

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“…Several manufacturers have used other variants such a "convergent chromatography" (Waters). Olesik [20] developed a variant where she only used 10-25% CO 2 and called the technique "LC with enhanced fluidity", or sometimes "enhanced fluidity chromatography".…”

Section: What Does "Sfc" Mean?mentioning

confidence: 99%

Instrumentation for analytical scale supercritical fluid chromatography

Berger

2015

Journal of Chromatography A

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“…Thus, by combining conventional SubFC with EFLC, the entire solvent range from 0–100% organic solvent is spanned. To date, EFLC has offered improved separations in reversed‐phase normal‐phase , size‐exclusion , chiral , and HILIC modes.…”

Section: Introductionmentioning

confidence: 99%

Enhanced‐fluidity liquid chromatography using mixed‐mode hydrophilic interaction liquid chromatography/strong cation‐exchange retention mechanisms

Beres

Olesik

2015

J of Separation Science

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The potential of enhanced-fluidity liquid chromatography, a subcritical chromatography technique, in mixed-mode hydrophilic interaction/strong cation-exchange separations is explored, using amino acids as analytes. The enhanced-fluidity liquid mobile phases were prepared by adding liquefied CO to methanol/water mixtures, which increases the diffusivity and decreases the viscosity of the mixture. The addition of CO to methanol/water mixtures resulted in increased retention of the more polar amino acids. The "optimized" chromatographic performance (achieving baseline resolution of all amino acids in the shortest amount of time) of these methanol/water/CO mixtures was compared to traditional acetonitrile/water and methanol/water liquid chromatography mobile phases. Methanol/water/CO mixtures offered higher efficiencies and resolution of the ten amino acids relative to the methanol/water mobile phase, and decreased the required isocratic separation time by a factor of two relative to the acetonitrile/water mobile phase. Large differences in selectivity were also observed between the enhanced-fluidity and traditional liquid mobile phases. A retention mechanism study was completed, that revealed the enhanced-fluidity mobile phase separation was governed by a mixed-mode retention mechanism of hydrophilic interaction/strong cation-exchange. On the other hand, separations with acetonitrile/water and methanol/water mobile phases were strongly governed by only one retention mechanism, either hydrophilic interaction or strong cation exchange, respectively.

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Enhanced fluidity liquid chromatography for hydrophilic interaction separation of nucleosides

Cited by 30 publications

References 26 publications

Characterization of enhanced-fluidity liquid hydrophilic interaction chromatography for the separation of nucleosides and nucleotides

Characterization of enhanced-fluidity liquid hydrophilic interaction chromatography for the separation of nucleosides and nucleotides

Instrumentation for analytical scale supercritical fluid chromatography

Enhanced‐fluidity liquid chromatography using mixed‐mode hydrophilic interaction liquid chromatography/strong cation‐exchange retention mechanisms

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