title: Comprehensive profiling of ribonucleosides modification by affinity zirconium oxide-silica composite monolithic column online solid-phase microextraction - Mass spectrometry analysis.
abstract: More than 140 modified ribonucleosides have been identified in RNA. Determination of endogenous modified ribonucleosides in biological fluids may serve as non-invasive disease diagnostic strategy. However, detection of the modified ribonucleosides in biological fluids is challenging, especially for the low abundant modified ribonucleosides due to the serious matrix interferences of biological fluids. Here, we developed a facile preparation strategy and successfully synthesized zirconium oxide-silica (ZrO2/SiO2) composite capillary monolithic column that exhibited excellent performance for the selective enrichment of cis-diol-containing compounds. Compared with the boronate-based affinity monolith, the ZrO2/SiO2 monolith showed ∼2 orders of magnitude higher extraction capacity and can be used under physiological pH (pH 6.5-7.5). Using the prepared ZrO2/SiO2 composite monolith as the trapping column and reversed-phase C18 column as the analytical column, we further established an online solid-phase microextraction (SPME) in combination with liquid chromatography-mass spectrometry (online SPME-LC-MS/MS) analysis for the comprehensive profiling of ribonucleosides modification in human urine. Our results showed that 68 cis-diol-containing ribosylated compounds were identified in human urine, which is, to the best of our knowledge, the highest numbers of cis-diol-containing compounds were determined in a single analysis. It is worth noting that four modified ribonucleosides were discovered in the human urine for the first time. In addition, the quantification results from the pooled urine samples showed that compared to healthy controls, the contents of sixteen ribose conjugates in the urine of gastric cancer, eleven in esophagus cancer and seven in lymphoma increased more than two folds. Among these ribose conjugates, four ribose conjugates increased more than two folds in both gastric cancer and esophagus cancer; three ribose conjugates increased more than two folds in both gastric cancer and lymphoma; one ribose conjugate increased more than two folds in both esophagus cancer and lymphoma. The developed analytical method provides a good platform to study the modified ribonucleosides in human body fluids.
authors: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), Department of Chemistry, Wuhan University, Wuhan 430072, China.
authors: Han-Peng
authors: HP
authors: Jiang
authors: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), Department of Chemistry, Wuhan University, Wuhan 430072, China.
authors: Jie-Mei
authors: JM
authors: Chu
authors: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), Department of Chemistry, Wuhan University, Wuhan 430072, China.
authors: Meng-Dan
authors: MD
authors: Lan
authors: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), Department of Chemistry, Wuhan University, Wuhan 430072, China.
authors: Ping
authors: P
authors: Liu
authors: Center for Gene Diagnosis, Zhongnan Hospital of Wuhan University, Wuhan 430071, China.
authors: Na
authors: N
authors: Yang
authors: Center for Gene Diagnosis, Zhongnan Hospital of Wuhan University, Wuhan 430071, China.
authors: Fang
authors: F
authors: Zheng
authors: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), Department of Chemistry, Wuhan University, Wuhan 430072, China. Electronic address: bfyuan@whu.edu.cn.
authors: Bi-Feng
authors: BF
authors: Yuan
authors: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), Department of Chemistry, Wuhan University, Wuhan 430072, China. Electronic address: yqfeng@whu.edu.cn.
authors: Yu-Qi
authors: YQ
authors: Feng
mesh_terms: D016022:Case-Control Studies; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009369:Neoplasms / Q000652:urine; D012263:Ribonucleosides / Q000737:chemistry* / Q000652:urine*; D012266:Ribose / Q000737:chemistry / Q000652:urine; D012822:Silicon Dioxide / Q000737:chemistry*; D052617:Solid Phase Microextraction / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D015040:Zirconium / Q000737:chemistry*
chemical_list: D012263:Ribonucleosides; D012266:Ribose; D012822:Silicon Dioxide; D015040:Zirconium; C028541:zirconium oxide
keywords: Cancer; Enrichment; Modified ribonucleoside; Online solid-phase microextraction; Zirconium oxide-silica composite monolith