title: Highly-sensitive detection of free advanced glycation end-products by liquid chromatography-electrospray ionization-tandem mass spectrometry with 2,4,6-trinitrobenzene sulfonate derivatization.
abstract: The common derivatization method of primary amino groups by 2,4,6-trinitrobenzene sulfonate (TNBS) was applied to the detection of free adducts of advanced glycation end-products (AGEs), in combination with liquid chromatography-tandem mass spectrometry-multiple reaction monitoring (LC-MS/MS-MRM). The proposed TNBS-MS method provided a surprisingly significant improvement of the detection of AGE-free adducts (e.g., by a factor of >1000 for methylglyoxal-derived hydroimidazolone) with a detection limit of 1.0 nM (10 fmol/injection volume), which was due to the high ionization efficiency of the derived trinitrophenol moiety and its hydrophobicity. With the aid of stable-isotope-labeled internal standard (MG-H1-d3), the convenient TNBS method that allowed the derivation of AGE-free adducts bearing amino groups under mild reaction conditions (30 mM, pH 8.5, 30 min, 30 °C) also permitted successive detection and quantification of five typical AGE-free adducts at nanomolar levels in rat plasma (50 μL) with high reproducibility (2-9% of RSD) and recovery (93-113%), using LC-MS/MS-MRM.
authors.affiliation: Division of Bioscience and Bioenvironmental Sciences, Faculty of Agriculture, Graduate School, Kyushu University, 6-10-1 Hakozaki, Fukuoka, Japan.
authors.forename: Chihiro
authors.initials: C
authors.lastname: Hashimoto
authors.forename: Yuri
authors.initials: Y
authors.lastname: Iwaihara
authors.forename: Si Jing
authors.initials: SJ
authors.lastname: Chen
authors.forename: Mitsuru
authors.initials: M
authors.lastname: Tanaka
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Watanabe
authors.forename: Toshiro
authors.initials: T
authors.lastname: Matsui
mesh_terms: D002851:Chromatography, High Pressure Liquid; D017127:Glycation End Products, Advanced / Q000032:analysis*; D015394:Molecular Structure; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry; D014302:Trinitrobenzenesulfonic Acid / Q000737:chemistry*
chemical_list: D017127:Glycation End Products, Advanced; D014302:Trinitrobenzenesulfonic Acid