Characterization of Nucleosides and Nucleobases in Natural Cordyceps by HILIC–ESI/TOF/MS and HILIC–ESI/MS

Abstract

A method combining hydrophilic interaction chromatography (HILIC) and electrospray ionization mass spectrometry (ESI-MS) was developed for the characterization and determination of natural Cordyceps. Separation was achieved on a Waters Xbridge Amide column with gradient elution. Identification of 15 target nucleosides and nucleobases was based on retention time, UV spectra and mass measurements of the protonated molecules ([M+H] + ) and main fragment ions (ESI-TOF/MS). Eight non-target compounds were tentatively identified by ESI-TOF/MS. The 15 target compounds were quantified by HILIC-ESI-MS/MS using time-programmed selective ion monitoring or multiple reaction monitoring in positive-ion mode under optimized mass conditions. This technique showed good linearity, repeatability and recovery. This approach was also successfully implemented in the analysis of nucleosides and nucleobases in 12 batches of natural Cordyceps samples that were collected from different regions in China. The developed HILIC-ESI-MS method exhibited clear advantages in identifying and determining highly polar bioactive components in Cordyceps, as well as their quality control.

GC-FID determination of nucleobases guanine, adenine, cytosine, and thymine from DNA by precolumn derivatization with isobutyl chloroformate

“…The results of the analysis agreed with the reported values for C by Xia et al (2011) and C, T, and A with Xia et al (2009) for human plasma samples. The reported values for plant DNA samples also agreed with Zhao et al (2003). Present study…”

Diverse Mechanisms of Sulfur Decoration in Bacterial tRNA and Their Cellular Functions

“…While addition of 10 mM sodium or ammonium acetate into the aqueous buffer at a pH ~4.5 enhances the separation and resolution of individual nucleosides, the chromatography parameters must be tailored to the individual needs of each study, focusing on complete resolution of the analytes of interest. This concern has spurred the application of hydrophilic interaction liquid chromatography (HILIC), in order to increase retention and resolution of nucleosides carrying polar modifications [91,141,142]. Though LC with UV-Visible detection has been an invaluable tool for analysis of tRNA modifications in years past, it is limited by sensitivity and the requirement of commercially available standards (Table 3) to validate retention times.…”

A Platform for Discovery and Quantification of Modified Ribonucleosides in RNA

“…As uridine and its modified derivatives often elute near the solvent front, alternatives to RP HPLC, such as hydrophilic interaction chromatography (HILIC), are useful to enhance retention and resolution of polar compounds (Alpert, 1990; Zhao et al, 2013). HILIC retains polar compounds through interaction with the hydrophilic solid phase enhanced by a high percentage organic mobile phase.…”