کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
10550150 967024 2005 10 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
LC-ESI-MS/MS determination of 4-hydroxy-trans-2-nonenal Michael adducts with cysteine and histidine-containing peptides as early markers of oxidative stress in excitable tissues
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
LC-ESI-MS/MS determination of 4-hydroxy-trans-2-nonenal Michael adducts with cysteine and histidine-containing peptides as early markers of oxidative stress in excitable tissues
چکیده انگلیسی
A sensitive, selective, specific and rapid liquid chromatographic-electrospray ionization tandem mass spectrometric assay was developed and validated for the simultaneous determination in skeletal muscle of the Michael adducts between 4-hydroxy-trans-2-nonenal (HNE), one of the most reactive lipid peroxidation-driven unsaturated aldehyde, and glutathione (GSH) and the endogenous histidine-containing dipeptides carnosine (CAR) and anserine (ANS), with the final aim to use conjugated adducts as specific and unequivocal markers of lipid peroxidation. Samples (skeletal muscle homogenates from male rats) were prepared by protein precipitation with 1 vol. of a HClO4 solution (4.2%; w/v) containing H-Tyr-His-OH as internal standard. The supernatant, diluted (1:1, v/v) in mobile phase, was separated on a Phenomenex Sinergy polar-RP column with a mobile phase of water-acetonitrile-heptafluorobutyric acid (9:1:0.01, v/v/v) at a flow rate of 0.2 ml/min, with a run time of 12 min. Detection was on a triple quadrupole mass spectrometer equipped with an ESI interface operating in positive ionization mode. The acquisitions were in multiple reaction monitoring (MRM) mode using the following precursor → product ion combinations: H-Tyr-His-OH (IS): m/z 319.2 → 156.5 + 301.6; GS-HNE: m/z 464.3 → 179.1 + 308.0; CAR-HNE: m/z 383.1 → 110.1 + 266.6; ANS-HNE: m/z 397.2 → 109.1 + 126.1. The method was validated over the concentration ranges 1.5-90 (GS-HNE) and 0.4-40 (CAR-HNE, ANS-HNE) nmoles/g wet tissue, and the LLOQ were 1.25 and 0.33 pmoles injected respectively. The intra- and inter-day precisions (CV%) were <7.38% (≤10.90% at the LLOQs); intra- and inter-assay accuracy (RE%) was within ± 7.0% for all the concentrations (≤18% at the LLOQs). The method was applied to quantitate peptide-HNE Michael adducts in rat skeletal muscles exposed to oxidative stress to endogenously generate HNE, and the results indicate that CAR-HNE can be considered as an early, specific and stable marker of lipid peroxidation in excitable tissues.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Chromatography B - Volume 827, Issue 1, 15 November 2005, Pages 109-118
نویسندگان
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