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.
Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), Department of Chemistry, Wuhan University , Wuhan, Hubei 430072, P.R. China.
Shao-Ting
ST
Wang
Wei
W
Huang
Wei
W
Lu
Bi-Feng
BF
Yuan
Yu-Qi
YQ
Feng
D002846:Chromatography, Affinity; D012263:Ribonucleosides / Q000032:analysis*; D052616:Solid Phase Extraction / Q000379:methods*; D014025:Titanium / Q000737:chemistry*
D012263:Ribonucleosides; C009495:titanium dioxide; D014025:Titanium