title: Evaluation of column hardware on liquid chromatography-mass spectrometry of phosphorylated compounds.
abstract: The influences of column hardware, such as chromatographic tubes and frits, on liquid chromatography-mass spectrometry (LC-MS) analysis of phosphorylated compounds were evaluated. The signal to noise ratio (S/N) and the intensity of flavin adenine dinucleotide (FAD) using a glass lined tube and polyethylene frit (GL-PE) column was approximately 170 and 90 times higher, respectively, than those using conventional stainless steel tube and stainless steel frit (S-S) column. In addition, the retention time of FAD using GL-PE column was the shortest compared to other columns. Interaction between phosphorylated compounds and metal ions in the flow path in the S-S column was stronger than that between them and the GL-PE column. Thus, the metal ions in the flow path in GL-PE column were low. Since the specific surface area of a pair of frits was 70 times larger than that of a chromatographic tube (150 mm×2.1 mm), the frits were found to have more effective improvement of the S/N as well as the intensity than the chromatographic tubes, when phosphorylated compounds were analyzed by LC-MS. When the evaluated phosphorylated compounds were analyzed by LC-MS(/MS) using a GL-PE column, the intensity and S/N were increased.
authors.affiliation: Department of Chemistry and Biomolecular Science, Faculty of Engineering, Gifu University, 1-1 Yanagido, Gifu 501-1193, Japan; Chemicals Evaluation and Research Institute, 1600 Shimotakano, Sugito-machi, Kitakatsushika-gun, Saitama 345-0043, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Sakamaki
authors.affiliation: Chemicals Evaluation and Research Institute, 1600 Shimotakano, Sugito-machi, Kitakatsushika-gun, Saitama 345-0043, Japan.
authors.forename: Takeharu
authors.initials: T
authors.lastname: Uchida
authors.affiliation: Department of Chemistry and Biomolecular Science, Faculty of Engineering, Gifu University, 1-1 Yanagido, Gifu 501-1193, Japan.
authors.forename: Lee Wah
authors.initials: LW
authors.lastname: Lim
authors.affiliation: Department of Chemistry and Biomolecular Science, Faculty of Engineering, Gifu University, 1-1 Yanagido, Gifu 501-1193, Japan. Electronic address: take-t@gifu-u.ac.jp.
authors.forename: Toyohide
authors.initials: T
authors.lastname: Takeuchi
mesh_terms: D002853:Chromatography, Liquid / Q000295:instrumentation* / Q000379:methods; D007477:Ions; D013058:Mass Spectrometry / Q000295:instrumentation* / Q000379:methods; D008670:Metals; D010748:Phosphopeptides / Q000032:analysis*; D059629:Signal-To-Noise Ratio
chemical_list: D007477:Ions; D008670:Metals; D010748:Phosphopeptides
keywords: Chromatographic tube; Frit; Intensity; LC/MS; Phosphorylated compounds; S/N