Highly-Sensitive Detection of Free Advanced Glycation End-Products by Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry with 2,4,6-Trinitrobenzene Sulfonate Derivatization
journal contribution
posted on 2013-05-07, 00:00 authored by Chihiro Hashimoto, Yuri Iwaihara, Si Jing Chen, Mitsuru Tanaka, Toshiaki Watanabe, Toshiro MatsuiThe 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.
