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Updated: May 26, 2026

Chemiluminescence-based Assays for Detection of Nitric Oxide and its Derivatives from Autoxidation and Nitrosated Compounds
Published on: February 16, 2022
¹⁸O-Labeled nitrous acid and nitrite: Synthesis, characterization, and oxyhemoglobin-catalyzed oxidation to
Anke Böhmer1, Anja Mitschke, Anna Reib
1Institute of Clinical Pharmacology, Hannover Medical School, 30625 Hannover, Germany.
Abstract:
We describe a simple laboratory method for specific labeling of nitrite with ¹⁸O for use in chemical and biochemical studies in the area of nitric oxide research. NaNO₂ (0.1 mmol) is diluted in H₂¹⁸O (45 μl) and acidified with HCl (1 μl, 5 M), and the solution is allowed to equilibrate. Subsequently, the sample is mixed by vortexing with ethyl acetate (500 μl), and the organic phase is dried over anhydrous Na₂SO(4). Ethyl acetate is evaporated to dryness, and the residue is reconstituted in phosphate-buffered saline. In human blood hemolysate, oxyhemoglobin (HbFe¹⁶O₂) was shown to oxidize N¹⁸O₂⁻ to ¹⁶ON¹⁸O₂⁻.
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