Highly-Sensitive Detection of Free Advanced Glycation End-Products by Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry with 2,4,6-Trinitrobenzene Sulfonate Derivatization

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• 作者：Chihiro Hashimoto ; Yuri Iwaihara ; Si Jing Chen ; Mitsuru Tanaka ; Toshiaki Watanabe ; Toshiro Matsui
• 刊名：Analytical Chemistry
• 出版年：2013
• 出版时间：May 7, 2013
• 年：2013
• 卷：85
• 期：9
• 页码：4289-4295
• 全文大小：376K
• 年卷期：v.85,no.9(May 7, 2013)
• ISSN：1520-6882

文摘

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鈥搕andem mass spectrometry-multiple reaction monitoring (LC鈥揗S/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 掳C) also permitted successive detection and quantification of five typical AGE-free adducts at nanomolar levels in rat plasma (50 渭L) with high reproducibility (2鈥?% of RSD) and recovery (93鈥?13%), using LC鈥揗S/MS-MRM.