On-line desalting-mass spectrometry system for the structural determination of hydrophilic metabolites, using a column switching technique and a volatile ion-pairing reagent.
A novel desalting method, using a column switching technique and a volatile ion-pairing reagent, pentadecafluorooctanoic acid, was developed. This system allows hydrophilic and cationic compounds in a nonvolatile buffer to be directly introduced into a mass spectrometer for structural elucidation. The desalting procedure consists of four steps: (1) the fractionation of a target compound from a separation column, (2) the removal of salts with pentadecafluorooctanoic acid on the trap column, (3) the desorption of the compound from the trap column, and (4) the re-equilibration of the trap column with a pentadecafluorooctanoic acid solution. In this procedure, we investigated the methods for optimizing the desalting and re-equilibration steps. Various amino acids, including branched chain amino acids, aromatic amino acids, basic amino acids and methionine, after separation with phosphate buffer on a cation-exchange column, were successively desalted by this method, and were observed as protonated ions by mass spectrometry. This desalting system could be useful for the structural elucidation of unknown hydrophilic compounds eluted by conventional high-performance liquid chromatography methods, such as ion-exchange chromatography, with mobile phases containing nonvolatile salts. As an example, we present the structural elucidation of unknown metabolites in bovine serum.
Yoshida
Hiroo
H
Institute of Life Sciences, Ajinomoto Co., Inc. 1-1 Suzuki-cho Kawasaki-ku, Kawasaki, Kanagawa, Japan.
Mizukoshi
Toshimi
T
Hirayama
Kazuo
K
Miyano
Hiroshi
H
D000596:Amino Acids / Q000302:isolation & purification; D000818:Animals; D002210:Caprylates / Q000737:chemistry; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D005466:Fluorocarbons / Q000737:chemistry; D007202:Indicators and Reagents; D013058:Mass Spectrometry / Q000379:methods*; D009862:Online Systems; D012492:Salts / Q000737:chemistry; D014466:Ultraviolet Rays
D000596:Amino Acids; D002210:Caprylates; D005466:Fluorocarbons; D007202:Indicators and Reagents; D012492:Salts; C023036:perfluorooctanoic acid