Hello everyone! I want to test if my synthesized compound inhibit reaction of lipid peroxidation. I have searched for protocols in published literature but aren't clear enough since specific time was not mentioned. I found methods where sodium linoleate and AAPH were used. Is that suitable method or can I determine inhibition by TBARS method? Thank you in advance
You may measure the lipid peroxidation product MDA (derivatization of MDA with 2,4 dinitrophenylhydrazine (DNPH) ) by HPLC method which is more efficient technique than TBARS method.
You may follow the attached article.
Hello
Your question is not clear enough.
If you want to assay antioxidant capacity of your compounds, you may do it by in vitro or in vivo method. For in vitro, you can do DPPH and ABTS. For in vivo method you may use a model animal (mouse, rat, zebrafish,...) and treat it with the compound and an oxidant agent (heavy metals, pesticides, hydrogen peroxide ...). Then measure TBARS levels. You must optimize the antioxidant and pro oxidant concentration and time needed to find their effects.
Have a look at this paper:
Article Beyond DPPH: Use of Fluorescence-Enabled Inhibited Autoxidat...
you can incorporate your compound into in vitro prepared liposomes and measure kinetics of the lipid peroxidation reaction by using a flourescent probe. I think this is what you're looking for.
For in vivo systems this is quite hard. There are some people using C11-Bodipy as a flourescent probe in cell culture or tissue biopsies. In this you can kind of monitor the progression of lipid peroxidation. If you are interested in measuring also lipid peroxidation products (i.e. ketones, aldehydes,..) and dont have the possibility to use mass spectrometry then you could also use a coumarin based probe such as CHH.
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