I am doing a research project on antioxidant activities of plant extracts. I am trying to synthesize peroxynitrite for the peroxynitrite free radical scavening assay and I have been unable to do so.
I used a method similar to the one described in the paper 'Antoxidant and free radical scavening activity of Spondias pinnata' written by Bibhabasu Haszra, Santanu Biswas and Nripendranath Mandal.
- 5ml of acidified (0.6 Hcl) 0.7M H2O2
- 5ml NaNo2
on an ice bath for 1 second.
We then added 5ml of ice-cold 1.2M NaOH and treated the solution with MnO2 (prewashed with 1.2M NaOH) to remove excess H2O2
This was then incubated -20 degrees celius overnight. It says to then collect the peroxynitrite from the top of frozen mixture. There seems like their might be a layer on top but it is not clealy clearly visible. I repeated the experiment and once again did not seem to work.
My supervisor is away on holidays so I am alone on this one for a while. If anybody has synthesised peroxynitrite before successfully some tips and suggestions would be greatly appreciated.
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