Quality peroxynitrite can be produced in any laboratory Romina Medeiros

Peroxynitrite is a reactive oxygen and nitrogen specie generated in vivo when nitric oxide reacts with superoxide, being a strong oxidant and nitrating agent of biomolecules. Its preparations are used frequently in biochemical reactions, being reported many methods for its synthesis. Most of them are potentially dangerous and require special and expensive equipment. In the present work we propose a simple, fast, low cost synthesis method, with high yield and low content of contaminants; based on Hugues and Nicklin 1968. The method requires hydrogen peroxide, sodium nitrite, hydrochloric acid and sodium hydroxide as reagents, and material commonly available in any laboratory. The remaining hydrogen peroxide was removed with manganese dioxide which was also prepared in house from potassium permanganate, ethanol and water. The concentration of peroxynitrite was determined spectrophotometrically, of hydrogen peroxide with a horseradish peroxidase-based assay, and of nitrite with the Griess method. Peroxynitrite was obtained with an approximate 80% yield, which decreased to approximately 70% after treatment with manganese dioxide, a hydrogen content lower than the assay detection limit (50 μM) and a nitrite concentration of less than 5%.