Analysis of nucleotides, nucleosides, nucleobases in cells by ion-pair reversed-phase HPLC

Simultaneous quantification of GMP, AMP, cyclic GMP and cyclic AMP by liquid chromatography coupled to tandem mass spectrometry

“…Inherent to their structure, the separation of nucleotides is usually performed by anion-exchange chromatography, though the mobile phase compositions are highly incompatible with LC-MS. Consistent with this study, the nucleotides already had been analyzed by reverse-phase high-performance liquid chromatography (RP-HPLC), with or without an ion-pairing reagent [19,21]. In this study, ionpairing reagents were not used because of their substantial background contribution and source pollution [22,23]; however, the nucleotides were eluted far from the column void volume although there was no difference in the retention times between nucleotides.…”

Development and application of an LC–MS/MS method for measuring the effect of (partial) agonists on cAMP accumulation in vitro

“…Another approach used is the separation of nucleotides using a reversed phase chromatography, however, because of their polar properties the nucleotides are poorly retained on a reversed stationary phase and therefore often used in combination with an ion-pairing reagent (N,Ndimethylhexylamine, Cai, 2001;Qian et al, 2004;hexylamine, Coulier et al, 2006, for review see Werner, 1991) or Hypercarb stationary phases combination with diethylamine (Xing et al, 2004). Preliminary experiments using the volatile ion-pairing reagent N,Ndimethylhexylamine (DMHA) showed a main peak of m/z 130 in our spectrum, which was originating from the DMHA, and resulted in poor sensitivity due to substantial background contribution, ion suppression, source pollution and possibly other side effects like space charging in the ion trap mass spectrometer.…”

Artemia enriched with hydrolyzed yeast improves growth and stress resistance of marine pejerrey Odontesthes argentinensis larvae

“…Prior to analysis, samples were freeze-dried. Nucleotides were separated and quantified by reversed-phase HPLC following the description of Werner (1991), and RNA was evaluated by the perchloric acid method (Herbert et al, 1971). …”