A novel gas chromatographic method for determination of malondialdehyde from oxidized DNA

Malondialdehyde (MA) is known to form from various lipids upon oxidation as one of secondary oxidation products. Determination of MA formed from lipid peroxidation has been used to examine occurrence of oxidative damages associated with many diseases, such as cancer, Alzheimer's, arthritis, inflammation, diabetes, atherosclerosis, and AIDS as well as aging. Analysis of MA is, however, extremely difficult because it is highly reactive and readily polymerized and forming adducts with biological substances such as proteins, phospholipids, and DNA (Shibamoto, J Pharm Biomed Anal 41:12-25, 2002). Gas chromatographic method using stable derivative, 1-methylpyrazole was advanced and has been successfully used to analyze MA in various lipids and lipid-rich foods. This method was also applied to determine MA formed from DNA and related compounds. The amounts found in oxidized 2'-deoxyribonucleotides were 213.8 nmol/16 mmol in 2'-deoxyguanosine, 130.6 nmol/16 mmol in 2'-deoxycytidine, 85.1 nmol/16 mmol in 2'-deoxyadenosine, and 84.5 nmol/16 mmol in thymidine. When the antioxidant activity of flavonoids and anthocyanins against calf thymus DNA oxidized with Fenton's reagent was examined using this newly developed gas chromatographic method, antioxidant activity of flavonoids and anthocyanins ranged from 48.5% (catechin) to 29.9% (apigenin) and from 45.0% (callistephin) to 10.2% (cyaniding), respectively.