We use a protocol based on the sample reaction with the thiobarbituric acid, using a kit of Cayman Chemical. 

As color reactive we use acetic acid and sodium hydroxide.

We built a standard curve with serial dilutions of malondialdehyde. 

@Maiara Providelo Thank you for you answer. But I don't  want to use Commercial Kit due to the high cost.

Thanks,

Éder

Take a look at the reference below. It describes a method for 4 different parameters (free and protein-bound MDA, lipid hydroperoxides and protein hydroperoxides) related to lipid peroxidation (including the one you need, MDA).

Grintzalis K, Zisimopoulos D, Grune T, et al (2013) Method for the simultaneous determination of free/protein malondialdehyde and lipid/protein hydroperoxides. Free Radic Biol Med 59:27–35. doi: 10.1016/j.freeradbiomed.2012.09.038

http://www.sciencedirect.com/science/article/pii/S0891584912011720

Dear Éder

I work on MDA for a long time, the problem with MDA assay that the reaction needs heating to boiling , that's make it ahrd to assay using microplate. but a new kit found using microplate but as you say it's expensive. 

Try preparing the TCA-TBA-HCl stock consisting of 15g trichloroacetic acid, 0.375g thiobarbituric acid in 100 mls of 0.25N HCl

Ekaette Fadairo Please is there any journal to back up this mixture preparation?

Yes Kunle. I got the SOP directly from the reagents dealer (PYREX) and I know that as long as you adhere strictly to the SOP within the right laboratory environmental conditions. the results come out well. The right SOP for your sample preparation should also be adhered to.