Highly-sensitive detection of free advanced glycation end-products by liquid chromatography-electrospray ionization-tandem mass spectrometry with 2,4,6-trinitrobenzene sulfonate derivatization

Chihiro Hashimoto, Yuri Iwaihara, Si Jing Chen, Mitsuru Tanaka, Toshiaki Watanabe, Toshiro Matsui

Research output: Contribution to journalArticle

24 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)4289-4295
Number of pages7
JournalAnalytical chemistry
Volume85
Issue number9
DOIs
Publication statusPublished - May 7 2013

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry

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