Separation of nucleosides and nucleobases

Being one of the basic components of all cells as building blocks of DNA and RNA, there is a high and growing need for bioanalytical methods to determine nucleosides, nucleobases and related compounds in a variety of samples. Hydrophilic interaction chromatography is one of the most suitable approaches for this, due to the hydrophilicity of these compounds as demonstrated by their negative logP and logD values.

This application shows baseline separation of the nucleosides, uridine, adenosine, and cytidine, together with their corresponding nucleobases, thymine, uracil, adenine, and cytosine, on a zwitterionic column with a bonded phosphorylcholine stationary phase under HILIC conditions that do not compromise the solubility of the compounds.

Isocratic separation of four nucleosides and their nucleobases on a Merck SeQuant ZIC-cHILIC column (100×4.6 mm) using an eluent containing 5 mM ammonium acetate in 80% acetonitrile/water, pumped at 0.5 mL/min at 22 °C. Eluent pumping, injection and detection by an Agilent HP 1050 system with quaternary pump, autosampler, and diode array detector (at 254 nm) controlled by ChemStation A10.01 software. Injection of 5 µL standards diluted in eluent to concentrations of 5-10 mg/L. Toluene was used as an approximate marker for the void volume.

Diduco is a leading provider of professional services in high performance liquid chromotography and producer of the Xenoic® XAMS chemical suppressor and ASUREX automatic regenerator for ion chromatography.

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