TiO2-Based Solid Phase Extraction Strategy for Highly Effective Elimination of Normal Ribonucleosides before Detection of 2′-Deoxynucleosides/Low-Abundance 2′-O-Modified Ribonucleosides
A novel TiO2-based SPE strategy was developed for eliminating normal ribonucleosides before mass spectrometry (MS) analysis of 2′-deoxynucleosides and 2′-O-modified ribonucleosides. The chromatographic research for the retention behavior of ribonucleosides and 2′-deoxynucleosides on TiO2 materials was investigated using TiO2 separation column. The results indicated a specific affinity interaction mechanism between TiO2 and cis-diol-containing ribonucleosides, and the interaction was proved effective even under a wide range of pH conditions and salt concentrations. Benefiting from these features, a TiO2-based solid phase extraction (SPE) method was developed for highly efficient elimination of RNA contamination from genomic DNA. Compared with the widely used enzymatic digestion method, the proposed TiO2-based SPE method showed much more efficiency for the removal of RNA as well as provided high recoveries for the 2′-deoxynucleosides. In addition, the sample processing time is dramatically shortened using the TiO2-based SPE method (∼5 min) compared to the traditional enzymatic digestion method (∼12 h). Finally, the purification of 2′-O-methylated ribonucleosides from RNA was successfully achieved in HeLa cells by the TiO2-based SPE method, which provided a proof-of-concept for the purification of relevant modified ribonucleosides from bulky normal ribonucleosides. Taken together, this strategy developed in the current study offers a promising option to purify 2′-deoxynucleosides/2′-O-modified ribonucleosides for their sensitive and accurate determination by eliminating normal ribonucleosides in biological samples.
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