Why going through this stress to assay for malondialdehyde? There is a simpler method, not requiring any standard curve.

MDA concentration, a marker of lipid peroxidation (LPO) can be determined by the method of Buege and Aust (1978). In this procedure, 0.1 mL of the serum or plasma or tissue homogenate (supernatant) will be added to 2 mL of trichloroacetic acid-thiobarbituric acid-hydrochloric acid (TCA/ TBA/HCl) (1:1:1 ratio) reagent, will be boiled at 100 degree C for 15 min, and allowed to cool. Flocculent materials will be removed by centrifugation at 3000 rpm for 10 min. The supernatant will be removed and the absorbance is read at 532 nm against blank. MDA concentration will be calculated using the molar extinction coefficient for MDA-TBA complex of 1.55 10^6 per M per cm.

In other words, you will only need 15% TCA, 0.385% TBA, and 0.25N HCl. The function of TCA is to precipitate any protein and other flocculent materials present. TBA will react with the corresponding MDA in your sample, and the reaction can only takes place in acidic medium (the reason for HCl). The pink color formed is equivalent to the concentration of MDA in sample. The thicker the pink color, the higher the MDA concentration and vice versa (Beer Lambert's principle).

I hope you will find it useful.

Thank you so much Oluwatobi T. Somade

You are welcome!

You can calculate the concentration using the formula:

C = A/Eb

Where c = concentration

A = absorbance

E = molar extinction coefficient (1.55 x 10^6)

b = pathlenght of 1cm