title: Gradient enhanced-fluidity liquid hydrophilic interaction chromatography of ribonucleic acid nucleosides and nucleotides: A "green" technique.
abstract: A "green" hydrophilic interaction liquid chromatography (HILIC) technique for separating the components of mixtures with a broad range of polarities is illustrated using enhanced-fluidity liquid mobile phases. Enhanced-fluidity liquid chromatography (EFLC) involves the addition of liquid CO2 to conventional liquid mobile phases. Decreased mobile phase viscosity and increased analyte diffusivity results when a liquefied gas is dissolved in common liquid mobile phases. The impact of CO2 addition to a methanol:water (MeOH:H2O) mobile phase was studied to optimize HILIC gradient conditions. For the first time a fast separation of 16 ribonucleic acid (RNA) nucleosides/nucleotides was achieved (16min) with greater than 1.3 resolution for all analyte pairs. By using a gradient, the analysis time was reduced by over 100% compared to similar separations conducted under isocratic conditions. The optimal separation using MeOH:H2O:CO2 mobile phases was compared to MeOH:H2O and acetonitrile:water (ACN:H2O) mobile phases. Based on chromatographic performance parameters (efficiency, resolution and speed of analysis) and an assessment of the environmental impact of the mobile phase mixtures, MeOH:H2O:CO2 mixtures are preferred over ACN:H2O or MeOH:H2O mobile phases for the separation of mixtures of RNA nucleosides and nucleotides.
authors: Department of Chemistry and Biochemistry, The Ohio State University, 100 West 18th Avenue, Columbus, OH 43210, United States.
authors: Michael C
authors: MC
authors: Beilke
authors: Department of Chemistry and Biochemistry, The Ohio State University, 100 West 18th Avenue, Columbus, OH 43210, United States.
authors: Martin J
authors: MJ
authors: Beres
authors: Department of Chemistry and Biochemistry, The Ohio State University, 100 West 18th Avenue, Columbus, OH 43210, United States. Electronic address: olesik.1@osu.edu.
authors: Susan V
authors: SV
authors: Olesik
mesh_terms: D000097:Acetonitriles; D002853:Chromatography, Liquid / Q000379:methods; D055772:Green Chemistry Technology; D057927:Hydrophobic and Hydrophilic Interactions; D000432:Methanol / Q000737:chemistry; D009705:Nucleosides / Q000032:analysis*; D009711:Nucleotides / Q000032:analysis*; D012313:RNA / Q000032:analysis*; D012997:Solvents; D014783:Viscosity; D014867:Water / Q000737:chemistry
chemical_list: D000097:Acetonitriles; D009705:Nucleosides; D009711:Nucleotides; D012997:Solvents; D014867:Water; D012313:RNA; D000432:Methanol; C032159:acetonitrile
keywords: Enhanced-fluidity liquid chromatography; Green chemistry; Hydrophilic interaction liquid chromatography; Nucleosides; Nucleotides