MDA by HPLC

Determination of free MDA by HPLC Below a pH of 4.65, free MDA exists in an undissociated cyclic form ( 245 = 30000). Although it can be determined by UV-absorptiometry (Kwon and Watts, 1963 Kukuda et al., 1981) and differential pulse polarography, HPLC techniques are far superior, because they are sensitive and specific. 

A number of HPLC methods for the detection of free MDA have been reported which include a size-exclusion method, a method that determined MDA-protein adducts, and a method that involves the determination of the MDA anion. 

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FIGURE 6. Inhibition of in vivo protein (carbonyls) and lipid peroxidation (MDA by HPLC) and of in vitro lipid peroxidation (TBARS-NADPH) in liver of guinea pigs fed during five weeks with high vitamin C diets versus those fed a diet just enough to avoid scurvy (33 mg ascorbic acid/kg). MDA = true malondialdehyde TEARS = thiobarbituric acid-reactive substances. TEARS were measured after 90 min in the presence of 0.2 mM FeS04, 5 mM ADP, and 1 mM NADPH. Reproduced with permission from Free Rad. Biol. Med. 17 105-115 (Barja et al., 1994). 

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