Stainless steel electrospray probe: a dead end for phosphorylated organic compounds?
A study of the interaction of phosphorylated organic compounds with the stainless components of a liquid chromatography-electrospray ionisation-mass spectrometry system (LC-ESI-MS) was carried out to disclose a (forgotten?) likely pitfall in the LC-ESI-MS analysis of phosphorylated compounds. The retention behaviour of some representative compounds of different important classes of phosphorylated biomolecules such as nucleotides, oligonucleotides, phosphopeptides, phospholipids and phosphorylated sugars was investigated during their passage through the injector and the stainless steel electrospray capillary. It became clear that the stainless steel components within the LC-ESI-MS setup were able to retain and trap phosphorylated compounds when these compounds were introduced under acidic conditions (0.1% acetic acid). Their release from these stainless steel parts was accomplished by applying an extreme basic mobile phase (25-50% ammonium hydroxide, ca. pH 12). From the data collected one could conclude that the availability of a primary phosphate group appeared imperative but was not always sufficient to realise adsorption on a stainless surface. Furthermore, the number of phosphate moieties seemed to enhance the adsorption properties of the molecules and hence roughly correlated with the analyte fraction lost. Corrosion of the inner surface caused by the mobile phase and the electrospray process was found to be an important factor in the course of these adsorption phenomena.
Tuytten
R
R
Department of Chemistry, Nucleoside Research and Mass Spectrometry Unit, University of Antwerp, Groenenborgerlaan 171, B-2020 Antwerp, Belgium.
Lemière
F
F
Witters
E
E
Van Dongen
W
W
Slegers
H
H
Newton
R P
RP
Van Onckelen
H
H
Esmans
E L
EL
D000327:Adsorption; D000595:Amino Acid Sequence; D008855:Microscopy, Electron, Scanning; D008969:Molecular Sequence Data; D009930:Organic Chemicals / Q000737:chemistry*; D010748:Phosphopeptides / Q000737:chemistry; D010766:Phosphorylation; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation*
D009930:Organic Chemicals; D010748:Phosphopeptides