A new way to fast and high resolution determination of modified nucleosides

The abnormal concentration of several modified nucleosides in the urine is supposed to be marker of carcinogenesis. For this reason analytical methods useful in the determination of these compounds in biological samples are of great importance. Present study concerns the application of Ultra High Performance Liquid Chromatography for fast, precise, high resolution separation and quantification of eight modified nucleosides in the synthetic serum and urine samples. The systematic study concerning the retention behavior of analyzed nucleosides on various stationary and mobile phases was performed. The attempt to apply four different column packings (octedecyl, octyl, phenyl, siliga gel) of different particle sizes and various mobile phases was made. On the basis of obtained results Kinetex C18 column and methanol/water mixtures were chosen for the utilization in biological samples. Developed method allows separation and quantification of eight modified nucleosides in serum or urine during 4 min with good linearity, accuracy and low LOQ values.

► UHPLC is a highly sensitive, specific and rapid technique for the analysis of modified nucleosides.
► Octadecyl UHPLC columns are the most appropriate for modified nucleosides analysis.
► Eight modified nucleosides were separated during 4 min in synthetic serum and urine.