Determination of Nucleosides in Escherichia coli by Rapid Resolution Liquid Chromatography–Tandem Quadrupole Mass Spectrometry

A sensitive and selective method was developed for the quantitative determination of nucleosides in Escherichia coli. An Agilent 1290 HPLC was used with mobile phase A (10 mM ammonium acetate) and mobile phase B (acetonitrile) at a flow-rate of 0.2 mL min−1 coupled with a mass spectrometry scanned in multiple reaction monitoring (MRM) mode. In the result, the limit of quantification and detection of 19 nucleosides were in the range of 0.05–20 μg L−1 and 0.02–10 μg L−1. The linearity of the detected nucleosides was excellent with R2 > 0.99 and the limits of detection were all satisfied. The recoveries for the compounds were between 79.0%–119.8%, and the relative standard deviation was below 14%. The method was capable of quantitation 7 nucleosides in Escherichia coli, of which guanosine was found to decrease along with culture phases.

T3 column provided 100% aqueous compatibility with bonding utilizes a high strength silica phase at a ligand density. 19 mixed nucleoside standards were analyzed by RRLC-MS/MS in MRM mode with T3 column, the nucleosides were readily separated in fourteen minutes, and the retention time of the majority were at 6–9 min.Figure optionsDownload as PowerPoint slide