Nitrosative deamination of 2′-deoxyguanosine and DNA by nitrite, and antinitrosating activity of β-carboline alkaloids and antioxidants

Endogenous and dietary nitrite produces reactive nitrogen species (RNS) that react with DNA causing mutations. The nitrosation of 2'-deoxyguanosine (dGuo) and DNA with nitrite was studied under different conditions, and the reaction and degradation products identified and analysed by HPLC-DAD-MS. Nitrosative deamination of dGuo produced xanthine along with 2'-deoxyxanthosine whereas DNA afforded xanthine. Formation of xanthine increased with nitrite concentration and in low pH such as that of stomach. Xanthine was measured as a marker of nitrosation of dGuo and DNA, and it was subsequently used to study the antinitrosating activity of beta-carboline alkaloids, and selected antioxidants. Food-occurring tetrahydro-beta-carbolines (TH beta Cs) decreased nitrosative deamination of dGuo and DNA under conditions simulating the stomach. Antinitrosating activity was also evidenced for flavonoids (catechin, quercetin) and indole (melatonin) antioxidants. Among TH beta Cs the most active antinitrosating compounds were 1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acids (TH beta C-3-COOHs) that reacted with nitrite to give N-nitroso derivatives as main products along with 3,4-dihydro-beta-carboline-3-carboxylic acids and aromatic beta-carbolines (norharman and hannan). Antinitrosating activity of TH beta Cs correlated well with the formation of N-nitroso-TH beta C-3-COOHs. These N-nitroso derivatives were stable at pH 7 but degraded in acid conditions affording nitrosating species.