High-throughput screening of the nucleosides and nucleotides using characteristic structural fragments fusion

Cordyceps sinensis is a traditional Chinese food. A high-throughput screening and quantitative method based on hydrophilic interaction chromatography coupled to quadrupole/Orbitrap high resolution mass spectrometry has been proposed for the joint measurement of nucleosides and their corresponding nucleotides in thirty-two Cordyceps sinensis samples. A total of thirty-two natural samples from six regions were enrolled. A multiple-step analysis strategy, including features extraction, noise level evaluation, mass shift correction and suspect spectral library spectra searching, was employed to discover, identify and validate the nucleosides and nucleotides. The remaining unknown compounds were identified with the characteristic structural fragments. The on-line dispersive solid-phase extraction system eliminates human-based steps errors and affords superior method reproducibility in the presence of hexafluoro-2-propanol and diethylamine as hydrophilic ion-pairing reagents. In view of the great difference of the quality, this high coverage confirmation strategy is of great reference value to the future quantitative analysis of potential nucleosides and nucleotides. Nucleosides and nucleotides ingredients could be considered as origins and quality assessment index of C. sinensis.