title: High-throughput LC-MS/MS based simultaneous determination of polyamines including N-acetylated forms in human saliva and the diagnostic approach to breast cancer patients.
abstract: The determination of polyamines and their N-acetylated forms was performed by ultraperformance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS). The polyamines efficiently reacted with 4-(N,N-dimethylaminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole (DBD-F) in 0.1 M borax (pH 9.3) at 60 °C for 30 min. The resulting derivatives were analyzed by electrospray ionization (ESI)-MS and sensitively detected by selected reaction monitoring (SRM). Furthermore, a rapid separation of the polyamine derivatives within 10 min was performed by UPLC using an antipressurized column packed with 1.7-μm octadecylsilyl (ODS) silica gel. The limits of detection (S/N = 3) on the SRM chromatograms were at the attomole level (9-43 amol). This procedure was used to successfully determine 11 polyamines, including their N-acetylated forms, in the saliva of patients with primary and relapsed breast cancer and healthy volunteers. The level of several polyamines (Ac-PUT, Ac-SPD, Ac-SPM, DAc-SPD, and DAc-SPM) increases in breast cancer patients. Furthermore, the levels of three polyamines (Ac-SPM, DAc-SPD, and DAc-SPM) were significantly higher only in the relapsed patients. The present method proved highly sensitive and is characterized by specificity and feasibility for sample analysis. Consequently, the proposed method is useful for the noninvasive salivary diagnosis of cancer patients and could be applied to determine polyamines in several specimens of biological nature.
authors.affiliation: Laboratory of Analytical and Bio-Analytical Chemistry, School of Pharmaceutical Sciences, University of Shizuoka , 52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan.
authors.forename: Haruhito
authors.initials: H
authors.lastname: Tsutsui
authors.forename: Toshiki
authors.initials: T
authors.lastname: Mochizuki
authors.forename: Koichi
authors.initials: K
authors.lastname: Inoue
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Toyama
authors.forename: Nobuyasu
authors.initials: N
authors.lastname: Yoshimoto
authors.forename: Yumi
authors.initials: Y
authors.lastname: Endo
authors.forename: Kenichiro
authors.initials: K
authors.lastname: Todoroki
authors.forename: Jun Zhe
authors.initials: JZ
authors.lastname: Min
authors.forename: Toshimasa
authors.initials: T
authors.lastname: Toyo'oka
mesh_terms: D000107:Acetylation; D001943:Breast Neoplasms / Q000175:diagnosis*; D016022:Case-Control Studies; D002853:Chromatography, Liquid / Q000379:methods*; D010080:Oxazoles / Q000737:chemistry; D011073:Polyamines / Q000032:analysis* / Q000737:chemistry; D012463:Saliva / Q000737:chemistry*; D013449:Sulfonamides / Q000737:chemistry; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013997:Time Factors
chemical_list: D010080:Oxazoles; D011073:Polyamines; D013449:Sulfonamides; C060548:4-(N,N-dimethylaminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole