The state-of-the-art determination of urinary nucleosides using chromatographic techniques “hyphenated” with advanced bioinformatic methods

Abstract

Over the last decade metabolomics has gained increasing popularity and significance in life sciences. Together with genomics, transcriptomics and proteomics, metabolomics provides additional information on specific reactions occurring in humans, allowing us to understand some of the metabolic pathways in pathological processes. Abnormal levels of such metabolites as nucleosides in the urine of cancer patients (abnormal in relation to the levels observed in healthy volunteers) seem to be an original potential diagnostic marker of carcinogenesis. However, the expectations regarding the diagnostic value of nucleosides may only be justified once an appropriate analytical procedure has been applied for their determination. The achievement of good specificity, sensitivity and reproducibility of the analysis depends on the right choice of the phases (e.g. sample pretreatment procedure), the analytical technique and the bioinformatic approach. Improving the techniques and methods applied implies greater interest in exploration of reliable diagnostic markers. This review covers the last 11 years of determination of urinary nucleosides conducted with the use of high-performance liquid chromatography in conjunction with various types of detection, sample pretreatment methods as well as bioinformatic data processing procedures.Electronic supplementary materialThe online version of this article (doi:10.1007/s00216-011-4789-6) contains supplementary material, which is available to authorized users.

Metabolomics in cell culture—A strategy to study crucial metabolic pathways in cancer development and the response to treatment

“…Comparison of supernatants from non-tumourigenic epithelial breast cell lines (MCF-10A) with early-stage breast cancer (MCF-7) cell lines has revealed remarkable differences in histidine metabolism, modified nucleosides, and metabolites derived from the polyamine/methionine cycle as well as compounds from purine de novo biosynthesis, which are all involved in the development of malignancy [71]. Besides, modified nucleosides, produced by the methylation and oxidative damage to the DNA [77], are not degraded or reutilised but distributed to the bloodstream and further eliminated with the urine [78]. Thus, presented molecules can be considered for further validation to extend the panel of metabolites already applied in urine samples to monitor breast cancer patients [79,80].…”

Gold-nanoparticle extraction and reversed-electrode-polarity stacking mode combined to enhance capillary electrophoresis sensitivity for conjugated nucleosides and oligonucleotides containing thioether linkers

“…Determination of normal and conjugate nucleosides and oligonucleotides in a variety of samples can be performed with different techniques, including immunoassays [5], highperformance liquid chromatography (HPLC), and capillary electrophoresis (CE) conjugated with UV detection [6,7] and mass spectrometry (MS) [8][9][10]. High-performance capillary electrophoresis (HPCE) has proved to be a rapid and simple technique for separating charged biomolecules and nanoparticles with very high resolution [11].…”

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

“…However low volatility of analyzed compounds as well as the need of their derivatization limits the application of this separation technique. Two interesting reviews were lately published dealing with the utilization of various separation techniques in the determination of modified nucleosides [16,17]. They concern mainly high performance liquid chromatography and electromigration techniques [16,17].…”