Profiling of cis-Diol-containing Nucleosides and Ribosylated Metabolites by Boronate-affinity Organic-silica Hybrid Monolithic Capillary Liquid Chromatography/Mass Spectrometry

Abstract

RNA contains a large number of modified nucleosides. In the metabolic re-exchange of RNA, modified nucleosides cannot be recycled and are thus excreted from cells into biological fluids. Determination of endogenous modified nucleosides in biological fluids may serve as non-invasive cancers diagnostic methods. Here we prepared boronate-affinity organic-silica hybrid capillary monolithic column (BOHCMC) that exhibited excellent selectivity toward the cis-diol-containing compounds. We then used the prepared BOHCMC as the on-line solid-phase microextraction (SPME) column and developed an on-line SPME-LC-MS/MS method to comprehensively profile cis-diol-containing nucleosides and ribosylated metabolites in human urine. Forty-five cis-diol-containing nucleosides and ribosylated metabolites were successfully identified in human urine. And five ribose conjugates, for the first time, were identified existence in human urine in the current study. Furthermore, the relative quantification suggested 4 cis-diol-containing compounds (5′-deoxy-5′-methylthioadensine, N4-acetylcytidine, 1-ribosyl-N-propionylhistamine and N2,N2,7-trimethylguanosine) increased more than 1.5 folds in all the 3 types of examined cancers (lung cancer, colorectal cancer, and nasopharyngeal cancer) compared to healthy controls. The on-line SPME-LC-MS/MS method demonstrates a promising method for the comprehensive profiling of cis-diol-containing ribose conjugates in human urines, which provides an efficient strategy for the identification and discovery of biomarkers and may be used for the screening of cancers.

Sensitive Determination of Onco-metabolites of D- and L-2-hydroxyglutarate Enantiomers by Chiral Derivatization Combined with Liquid Chromatography/Mass Spectrometry Analysis

“…Pretreatment of urine samples was performed according to the previous reported procedure 34 35 . Briefly, the freshly collected urine samples were immediately centrifuged at 5000 × g for 10 min under 4 °C twice to remove insoluble debris.…”

DNA hydroxymethylation age of human blood determined by capillary hydrophilic-interaction liquid chromatography/mass spectrometry

“…Recently, our group successfully developed a novel method for one-pot preparation of organic-silica hybrid capillary monolithic column by sol–gel combined with “thiol-ene” click reaction, which allows higher yield and less by-products compared to other methods [ 30 – 32 ]. Using the “thiol-ene” click reaction, here we successfully prepared hydrophilic organic-silica hybrid monolith.…”

Efficient and selective adsorption of small polyols by boronic acid functionalized polystyrene adsorbent

“…23,24 The selectivity and reversibility make boronic acid a good construction moiety for polymers for diol sensors, [25][26][27] drug delivery 28,29 and biomolecule enrichment. [30][31][32] Research on boronic acid modified polymeric materials for the separation of diol-containing biomolecules, such as nucleosides, [33][34][35] glycoprotein and glycopeptides, 36,37 have made good progress, but most of these materials are only suitable for small-scale separation of large biomolecules. Boronic acid functionalized adsorbents for the enrichment of short chain polyols which are capable of field application remain a research blank.…”