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Integration of Miniaturized Solid Phase Extraction and LC-MS/MS Detection of 3-Nitrotyrosine in Human Urine for Clinical Applications
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Comparison of spectrophotometric, HPLC and chemiluminescence methods for 3-nitrotyrosine and peroxynitrite
Nilufer Nermin Turan1, Mustafa Ark, Abdullah Tuncay Demiryurek
1Gazi University, Faculty of Pharmacy, Department of Pharmacology, 06330 Etiler, Ankara, Turkey. niluferturan@gazi.edu.tr
Abstract:
We have studied the interaction of 3-nitrotyrosine with peroxynitrite using three different methods; chemiluminescence, spectrophotometry and HPLC. Peroxynitrite-induced luminol or lucigenin chemiluminescence were significantly decreased by 3-nitrotyrosine, in concentration-dependent manners. The intensity of the peroxynitrite spectrum was also markedly reduced in the presence of 3-nitrotyrosine in the spectrophometric assay. However, there was no attenuation of the 3-nitrotyrosine signal in the HPLC assay after mixing with peroxynitrite. The interaction of 3-nitrotyrosine and hypochlorous acid (HOCl) was also studied via the chemiluminescence assay, where the HOCl-induced responses were markedly inhibited by 3-nitrotyrosine. These results suggest that caution should be taken when studying the levels or interactions of 3-nitrotyrosine.
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