Malondialdehyde measurement in urine

Abstract

Malondialdehyde (MDA) is an end product of lipid peroxidation and is a frequently measured index of these processes. The thiobarbituric acid (TBA) test is commonly used to measured MDA, but its specificity is questionable due to the presence of interfering chromogens. Wade and van Rij described in 1988 a method which removes these chromogens by HPLC. However, the sensitivity and the resolution of this method was not adequate for measurements of MDA in urine. We have improved this method by replacing TBA with diethylthiobarbituric acid (DETBA). The less polar MDA-DETBA complexes were isolated on Bakerbond cartridges and quantified by HPLC without interference. MDA was detectable using a fluorescence or ultraviolet detector at picomole levels. This technique was applied to urine samples obtained from ten burns patients on different days following their hospitalization. Urinary MDA in burns patients was very high and reached 18.6 mumol/mmol creatinine in one patient compared with a mean value of 0.23 mumol/mmol creatinine in healthy controls. Maximum MDA levels were attained on the third day for the majority of patients and remained, on average, much higher than normal even after 20 days. Using this method, picomole quantities of MDA can be easily and specifically detected in urine samples. This method is useful for assessing an oxidative stress.