Determination of Nucleosides in Escherichia coli by Rapid Resolution Liquid Chromatography–Tandem Quadrupole Mass Spectrometry
A sensitive and selective method was developed for quantitation of nucleosides. The method included a UPLC HSS T3 column separation operated with a mobile phase of 10 mmol/L ammonium acetate and acetonitrile at a flow-rate of 0.2 mL/min coupled to a mass spectrometry scanned in multiple reaction monitoring (MRM) mode. In the result, the limits of quantification and detection of 19 nucleosides were in the range of 0.05-20 mu g/L and 0.02-10 mu g/L, respectively. The linearity of the detected nucleosides was excellent with R-2 > 0.99 and the limits of detection were all satisfied. The recoveries of standard additions for the compounds were between 79.01% and 119.76%, and the relative standard deviation was below 14%. The method was capable of quantitation for 7 nucleosides in Escherichia coli, of which guanosine was found to decrease along with culture phases.